Formulations from natural products, turmeric, paclitaxel, and aspirin

ABSTRACT

The invention provides novel compounds and formulations of turmeric oil, fish oil, aspirin and anti-cancer drugs (paclitaxel) having anti-inflammatory, analgesic and/or anti-cancer activity.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. Ser. No.13/537,814, filed on Jun. 29, 2012, which claims benefit under 35 U.S.C.§120 and is a Continuation-in-Part of International PCT Application No.PCT/US2010/062481, filed Dec. 30, 2010, which claims benefit under 35U.S.C. §119(e) to U.S. Provisional Application Ser. No. 61/282,211,filed Dec. 31, 2009. Contents of both applications are incorporated byreference herein in their entireties.

FIELD OF THE INVENTION

The invention relates to compositions and methods for treatment ofinflammatiory diseases and cancer.

BACKGROUND OF THE INVENTION

Turmeric is an Asian spice and a traditional remedy since 600 BC. Theturmeric plant (Curcuma longa) is a member of the ginger family(Zingiberaceae). The rhizome (underground stem) is used to obtainturmeric powder or ground turmeric. Turmeric was used for many centuriesin Ayurveda, an Indian traditional medical system, for treating andpreventing a number of illnesses. Modern scientific studies of thepowerful biologically active compounds contained in the natural product,turmeric has been shown to have many health benefits including stronganti-cancer properties. It decreases symptoms of skin cancers andreduces the incidence of chemically caused breast cancer in animals.

Curcumin is an active ingredient derived from turmeric and it impartsthe yellow color in turmeric. It has several biological activities withbeneficial effects on cancer prevention and cure, and on a variety ofother diseases such as arthritis, wound healing and Alzheimer's disease.In addition to curcumin there are several other biologically activecompounds in turmeric which have not received much attention but havestrong biological activity. Curcumin has been shown to be effective inthree ways in attacking cancer. It suppresses transformation,proliferation, and metastasis of tumors. Curcumin has been shown to haveprotective and therapeutic effects against cancers of the blood, skin,pancreas, lung, oral cavity, and intestinal tract. Curcumin is shown tobe multi-targeted since it modulates multiple cell signaling pathways.Curcumin asserts its anti-tumor activity by altering the dysregulatedcell cycle via (a) cyclin-dependent, (b) p53-dependent and (c)p53-independent pathways.

However, curcumin is poorly bio-available following oral administration.The results suggest that doses of curcumin required to furnish hepaticlevels sufficient to exert pharmacological activity are probably notfeasible in humans and further research on both the biological activityand bioavailability of dietary polyphenols is needed to properly assesstheir usefulness for the prevention and treatment of disease.

SUMMARY OF THE INVENTION

The inventor has discovered a series of novel compounds and formulationshaving anti-inflammatory, analgesic and/or anti-cancer activity.Accordingly, in one aspect, the invention provides a turmeric oilextract obtained by high vacuum distillation of turmeric oil andcollecting a distillate at 70-100° C., at 105 to 118° C. or at 100-130°C.

In another aspect, the invention provides curcumin derivatives havingthe structure shown in formula (I):

wherein:

R¹ and R² are independently H, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted cyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, a peptide, —C(O)R³,—C(O)OR³, or —C(O)NR³R³, provided that at least one of R¹ and R² is notH, or both of R¹ and R² are not octyl or hexadecyl, or one of R¹ or R²is not H and the other is not octyl or hexadecyl;

R³ is independently for each occurrence H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl; and

analogs, derivatives, isomers, prodrugs, and pharmaceutically acceptablesalts thereof.

The inventor has also discovered that carbohydrate-aspirin conjugatesunexpectedly have enhanced anti-inflammatory and/or anti-cancer activityrelative to unconjugated aspirin. Thus, the invention provides compoundsof formula (II) as anti-inflammatory and anti-cancer agents. A compoundof formula (II) is:

wherein:

R⁸ is a carbohydrate;

R⁹ is H, optionally substituted alkyl, optionally substituted alkenyl,optionally substituted alkynyl, or optionally substituted acyl; and

analogs, derivatives, isomers, prodrugs, and pharmaceutically acceptablesalts thereof.

In yet another aspect, the invention provides a composition comprisingturmeric oil or a turmeric oil extract and at least one compoundselected from the group consisting of anti-cancer agents,anti-inflammatory agents, compounds of formula (I), compounds of formula(II), fish oil, fish oil extract, and any combinations thereof.

The invention also provides a composition comprising curcumin and/or acurcumin derivative of formula (I) and at least one compound selectedfrom the group consisting of anti-cancer agents, anti-inflammatoryagents, compounds of formula (II), fish oil, fish oil extract, and anycombinations thereof.

The invention further provides a method for treating a subject forinflammation, or a disease or condition associated with inflammation,the method comprising administering a therapeutically effective amountof a turmeric oil extract described herein, a compound described herein,a composition described herein, and any combinations thereof to asubject in need thereof.

The invention also provides a method of treating a subject for cancer,the method comprising administering a therapeutically effective amountof a turmeric oil extract described herein, a compound described herein,a composition described herein, and any combinations thereof to asubject in need thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1B depict exemplary curcumin derivatives of formula (I).

FIGS. 2A and 2B show the NMR spectra of turmeric oil extract fractionNJ-78-12. FIG. 2A shows the full spectra, and FIG. 2B shows the peaksfrom 5.2 ppm to 6.4 ppm.

FIGS. 3A-3C show the NMR spectra of turmeric oil extract fractionMT-133-3. FIG. 3A shows the full spectra; FIG. 3B shows peaks from 0 ppmto 2.6 ppm; FIG. 3C shows the peaks from 4.7 ppm to 6.3 ppm.

FIGS. 4A and 4B are HPLC spectra of NJ-78-12 (FIG. 4A) and MT-133-3(FIG. 4B).

FIG. 5 is a bar graph showing tail flick latency (seconds) in aspirinand glucose-aspirin conjugated treated animals (n=8 in each group).Group 1=aspirin 100 mg/kg; Group 2=aspirin 200 mg/kg; Group3=glucose-aspirin conjugate 100 mg/kg; Group 4=glucose-aspirin conjugate200 mg/kg; and Group 5=control.

FIG. 6 is a line graph showing percentage of maximal possible effect (%MPE) in various groups (n=8 in each group). Groups are same as in FIG.5.

FIG. 7 is a line graph showing paw edema circumference (cm) in variousgroups. Groups are same as in FIG. 5.

FIG. 8 is a bar graph showing percentage inhibition of paw edema invarious groups (n=8 in each group). Groups are same as in FIG. 5.

FIG. 9 is a bar graph showing percentage inhibition of paw edema withcontrols. Group 1=DMSO; Group 2=aspirin 100 mg/kg; Group 3=aspirin 200mg/kg; Group 4=normal saline. *P<0.05 as compared to Group 1; #P<0.05 ascompared to Group 4 is a bar graph showing percentage inhibition of pawedema with controls.

FIG. 10 is a bar graph showing percentage inhibition of paw edema withNJ1 (turmeric oil distillate 115° C.-120° C./high vacuum (˜100 torr).Groups are as shown in Table 4.

FIG. 11 is a bar graph showing percentage inhibition of paw edema withNJ2 (omega-3 DHA/EPA-fish oil). Groups are as shown in Table 4.

FIG. 12 is a bar graph showing percentage inhibition of paw edema withNJ3 (1:1 mix of turmeric oil distillate 115° C.-120° C./high vacuum(˜100 torr)+omega-3 DHA/EPA-fish oil). Groups are as shown in Table 4.

FIG. 13 is a bar graph showing percentage MPE in tail flick test withcontrols. Group 1=DMSO; Group 2=aspirin 100 mg/kg; Group 3=aspirin 200mg/kg; and Group 4=normal saline. *P<0.05 as compared to Group 1; #P<0.05 as compared to Group 4.

FIG. 14 is a bar graph showing percentage MPE in tail flick test withNJ1 (turmeric oil distillate 115° C.-120° C./high vacuum (˜100 torr).Groups are as shown in Table 4.

FIG. 15 is a bar graph showing percentage MPE in tail flick test withNJ2 (omega-3 DHA/EPA-fish oil). Groups are as shown in Table 4.

FIG. 16 is a bar graph showing percentage MPE in tail flick test withNJ3 (1:1 mix of turmeric oil distillate 115° C.-120° C./high vacuum(˜100 torr)+omega-3 DHA/EPA-fish oil). Groups are as shown in Table 4.

FIGS. 17A and 17B are bar graph (FIG. 17A) and line graph (FIG. 17B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with NJ-58-1 (diacetylcurcumin)at 48 hours. Non-cancer cell line WI-38 was used as a control. In FIG.17A, concentration is micromolar (μM), and in FIG. 17B concentration ismolar (M).

FIGS. 18A and 18B are bar graph (FIG. 18A) and line graph (FIG. 18B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with MT-76-1(curcumin-glutarate) at 48 hours. Non-cancer cell line WI-38 was used asa control. In FIG. 18A, concentration is micromolar (μM), and in FIG.18B concentration is molar (M).

FIGS. 19A and 19B are bar graph (FIG. 19A) and line graph (FIG. 19B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with curcumin at 48 hours.Non-cancer cell line WI-38 was used as a control. In FIG. 19A,concentration is micromolar (μM), and in FIG. 19B concentration is molar(M).

FIGS. 20A and 20B are bar graph (FIG. 20A) and line graph (FIG. 20B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with NJ-78-12 (turmeric oildistillate) at 48 hours. Non-cancer cell line WI-38 was used as acontrol.

FIGS. 21A and 21B are bar graph (FIG. 21A) and line graph (FIG. 21B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with MT-133-3 (turmeric oildistillate fraction). Non-cancer cell line WI-38 was used as a control.Cells were treated with MT-133-3 for 48 hours.

FIGS. 22A and 22B are bar graph (FIG. 22A) and line graph (FIG. 22B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with PMN 11-168(glucose-aspirin) at 48 hours. Non-cancer cell line WI-38 was used as acontrol. In FIGS. 22A and 22B concentration of compounds is in molar(M).

FIGS. 23A and 23B are bar graph (FIG. 23A) and line graph (FIG. 23B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with aspirin at 48 hours.Non-cancer cell line WI-38 was used as a control. In FIGS. 23A and 23Bconcentration of compounds is in molar (M).

FIGS. 24A and 24B are bar graph (FIG. 24A) and line graph (FIG. 24B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with NJ-81-4 (mixture ofturmeric oil distillation fraction NJ-78-12 (10 mg) and Paclitaxel(taxol) (1 mg)) at 48 hours. Non-cancer cell line WI-38 was used as acontrol. In FIGS. 24A and 24B concentration is in molar (M) with respectto paclitaxel.

FIGS. 25A and 25B are bar graph (FIG. 25A) and line graph (FIG. 25B)showing % cellular inhibition of various cell lines breast (SKBR3),pancreatic (Panc1), and prostate (PC-3) with taxol at 48 hours.Non-cancer cell line WI-38 was used as a control. In FIGS. 25A and 25Bconcentration is in molar (M).

FIGS. 26A-26D are line graphs showing inhibition rate of NJ-92-1(gemcitabine), NJ-92-2 (MT-133-3), NJ-92-3 (gemcitabine (1 mg)+MT-133-3(10 mg)), NJ-92-4 (Paclitaxel), and NJ-92-5 (Paclitaxel (1 mg)+MT-133-3(10 mg)) on cancer cell lines PANC-1 (FIG. 26A), PC3 (FIG. 26B) andSK-BR-3 (FIG. 26C). Non-cancer cell line WI38 (FIG. 26D) was used as acontrol. No LD₅₀ (lethal dose 50%) was observed for the testedcompounds; thus, demonstrating that these compounds were not cytotoxicunder the testing conditions.

FIGS. 27A-27D are line graphs showing inhibition rate of NJ-92-3 2K(2,000 ng/ml) (gemcitabine (1 mg)+MT-133-3 (10 mg)), NJ-92-3 20 (20ng/ml), NJ-92-5 2K (2000 ng/ml) (Paclitaxel (1 mg)+MT-133-3 (10 mg)),NJ-92-5 20 (20 ng/ml), Taxol 2K (2000 ng/ml), Taxol 20 (20 ng/ml), andDMSO on cancer cell lines PANC-1 (FIG. 27A), PC3 (FIG. 27B) and SK-BR-3(FIG. 27C). Non-cancer cell line WI38 (FIG. 27D) was used as a control.

FIGS. 28A-28D are line graphs showing inhibition rate of NJ-92-1(gemcitabine), NJ-92-2 (MT-133-3, NJ-92-3 (gemcitabine (1 mg)+MT-133-3(10 mg)), NJ-92-4 (Paclitaxel), and NJ-92-5 (Paclitaxel (1 mg)+MT-133-3(10 mg)) on cancer cell lines PANC-1 (FIG. 28A), PC3 (FIG. 28B) andSK-BR-3 (FIG. 28C). Non-cancer cell line WI38 (FIG. 28D) was used as acontrol.

FIGS. 29A and 29B show HPLC spectra of turmeric oil extracts NJ-78-12(FIG. 29A) and MT-133-3 (FIG. 29B). HPLC conditions: X-Terra C-18 column(4.6×150 mm, 5 μm), acetonitrile/water (85/15), 1 mL/min, UV detector @254 nm.

FIG. 30 is a bar graph showing % inhibition of paw edema.

FIG. 31 is a bar graph showing % MPE in tail flick test with controls.*P<0.05 as compared to Group 2.

FIGS. 32-38 show the NMR spectra of turmeric oil extract fractionsNJ-106-1 (FIG. 32), NJ-106-1 (FIG. 33), MT-133-1 (FIG. 34), MT-133-3(FIG. 35), MT-133-4 (FIG. 36), MT-133-5 (FIG. 37), and MT-133-8 (FIG.38).

FIG. 39 is a bar graph showing % inhibition of paw edema.

FIG. 40 is a bar graph showing % M PE in tail flick test with controls.MPE: P<0.05 as compared to control, at 60 min-with 4, 5, 6; at 90min-with 6; at 120 min-with 6; at 180 min-with 6.

FIGS. 41-43 show GC mass spectra of turmeric oil extract fractionBR-110-6.

FIGS. 44A and 44B show the NMR spectra of the two fractions obtainedfrom the turmeric oil extract fraction BR-110-4 after passing through asilica gel column and eluting with ethyl acetate and hexane.

FIGS. 45-48 show GC mass spectra of turmeric oil extract fractionNJ-100-9.

FIG. 49 shows structures of some exemplary sesquiterpene component s ofturmeric oil fractions. Show are ar-curcumene (1), 7-epizingiberene (2),β-sesquiphellandrene (3), β-bisabolene (4), curlone (5), α-turmerone(6), β-turmerone (7), and ar-turmerone (8).

DETAILED DESCRIPTION OF THE INVENTION

The inventor has discovered a series of novel compounds and formulationshaving anti-inflammatory, analgesic and/or anti-cancer activity.

Turmeric Oil Extract

In one aspect, described herein is a turmeric oil extract obtained byhigh vacuum distillation of turmeric oil and collecting a distillate at70-100° C., at 105 to 118° C. or at 100-130° C. As used herein, the term“high vacuum” refers to pressure less than about 250 torr of basevacuum. In some embodiments, distillation is at a pressure less thanabout 200 torr, less than about 150 torr, less than about 100 torr, lessthan about 50 torr, less than about 25 torr, less than about 10 torr,less than about 0.1 torr, or less than about 0.01 torr. In oneembodiment, distillation is at pressure in the range of 0.1 torr to 100torr.

In some embodiments the extract is obtained by a method comprising thesteps of: (i) extracting a turmeric powder with hexane; (ii) distillingthe extract of (i) to obtain a distillate at 115-135° C. under highvacuum; (iii) distilling the distillate of (ii) to obtain a distillateat 95-112° C. under high vacuum; (iv) distilling the distillate of (iii)to obtain a distillate at 100-110° C. under high vacuum; and (v)distilling the distillate of (iv) to obtain the extract as a distillateat 120-123° C. under high vacuum. This extract is also referred to asNJ-78-12 herein. It is to be understood that a distillate of (ii),(iii), or (iv) is also a turmeric oil extract of the invention.

In some embodiments the extract is obtained by a method comprising thesteps of: (i) extracting a turmeric powder with hexane; (ii) distillingthe extract of (i) to obtain a distillate at 115-135° C. under highvacuum; (iii) distilling the distillate of (ii) to obtain a distillateat 95-112° C. under high vacuum; (iv) distilling the distillate of (iii)to obtain a distillate at 100-110° C. under high vacuum; (v) distillingthe distillate of (iv) to obtain a distillate at 100-120° C. and at 124°C. under high vacuum; (vi) combining the distillates obtained in (v) andobtaining the extract by eluting the combined distillates from a columnusing one volume of hexane, one volume 0.5% of ethyl acetate/hexane, andhalf volume 1% ethyl acetate/Hexane. This fraction is also referred toas MT-133-3 herein.

Further fractions from the column can also be obtained as follows: afterelution of fraction MT-133-3, eluting with half volume of 1% ethylacetate/Hexane to obtain the fraction referred to as MT-133-4; followedby two volumes of 2% ethyl acetate/Hexane to obtain fractions referredto as MT-133-5, MT-133-6 and MT-133-7; followed by one volume of 5%ethyl acetate/hexane to obtain fractions MT-133-8 and MT-133-9; followedby ¼ volume of methanol to obtain fraction MT-133-10. It is to beunderstood that a distillate of (ii), (iii), (iv) or (v) is also aturmeric oil extract of the invention.

Furthermore, any of the fractions obtained from the columnchromatography purification, either alone or in a mixture with one ormore of the other fractions is also considered a turmeric oil extract ofthe invention.

In some embodiments the extract is obtained by a method comprising thesteps of: (i) extracting a turmeric powder with hexane; (ii) distillingthe extract of (i) to obtain a first distillate at below 108° C. undervacuum, a second distillate at 108° C.-122° C. under vacuum, and a thirddistillate at 122° C.-143° C. under vacuum; (iii) purifying the seconddistillate from (ii) using flash column chromatography withhexane-ethylacetate. This extract is also referred to as BR-110-6herein.

In some embodiments the extract is obtained by a method comprising thesteps of: (i) extracting a turmeric powder with hexane; (ii) distillingthe extract of (i) to obtain a first distillate at below 108° C. undervacuum, a second distillate at 108° C.-122° C. under vacuum, and a thirddistillate at 122° C.-143° C. under vacuum; (iii) purifying the seconddistillate from (ii) using flash column chromatography withhexane-ethylacetate; (iv) combining the purified product from (iii)(e.g., BR-110-6) with the third distillate of (ii); and (v) distillingthe combined mixture of (iv) to obtain a distillate at 118° C.-137° C.under vacuum. This distillate is also referred to as BR-132-4 herein andcorresponds to fractions MT-133-4 to MT-133-9 described herein.

In some embodiments the extract is obtained by a method comprising thesteps of: (i) extracting a turmeric powder with hexane; (ii) distillingthe extract of (i) to obtain a first distillate at below 108° C. undervacuum, a second distillate at 108° C.-122° C. under vacuum, and a thirddistillate at 122° C.-143° C. under vacuum; (iii) purifying the seconddistillate from (ii) using flash column chromatography withhexane-ethylacetate; (iv) combining the purified product from (iii)(e.g., BR-110-6) with the third distillate of (ii); (v) distilling thecombined mixture of (iv) to obtain a distillate at 118° C.-137° C. undervacuum. This distillate is also referred to as BR-132-4 herein andcorresponds to fractions MT-133-4 to MT-133-9 described herein; and (vi)purifying the distillate of (v) to obtain two fractions. These purifiedfractions are also referred to as NJ-106-1 and NJ-106-2 herein.

In some embodiments the extract is obtained by a method comprising thesteps of: (i) extracting a turmeric powder with hexane; (ii) distillingthe extract of (i) to obtain a distillate at 115° C.-135° C.; (iii)distilling the distillate of (ii) to obtain a first distillate at 95°C.-108° C. under vacuum and a second distillate at 108° C.-112° C.; (iv)distilling the second distillate from (iii) to obtain a first distillateat 100° C.-120° C., a second distillate at 120° C.-123° C., and a thirddistillate at 124° C. under high vacuum. These turmeric oil extracts arealso referred to as NJ-78-11 (first distillate), NJ-78-12 (seconddistillate), and NJ-78-13 (third distillate) herein.

In some embodiments, the method of obtaining the turmeric oil extractfurther comprises the step of purifying the extract obtained by theabove methods. The skilled artisan is well aware of methods of purifyingcompounds. Such methods include, but are not limited to, columnchromatography, high pressure liquid chromatography, size-exclusionchromatography, crystallization, distillation, filtration, and the like.

The turmeric oil fraction of the invention may contain one or moresesquiterpenes including, but not limited to, Ar-turmerone α-turmerone,and β-turmerone.

In some embodiments, the turmeric oil fraction comprises a mixture ofcompounds with at least one compound comprising at least 50%, at least60%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, or at least 95% of the mixture of the compounds. The amount of thecompounds in the turmeric oil fraction can be determined using anytechniques available to the skilled artisan, including, but not limitedto, high performance liquid chromatography (HPLC), liquidchromatography, size exclusion chromatography, thin layer chromatography(TLC), NMR, and IR.

In some embodiments, the turmeric oil extract is at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least95% pure as determined by NMR, HPLC, LC and/or TLC. In some embodiments,the purity of the turmeric oil extract is determined by HPLC with thefollowing conditions: X-Terra C-18 column (4.6×150 mm, 5 μm),Acetonitrile/water (85/15), 1 mL/min, UV detection at 253 nm.

The turmeric oil extract of the invention can be identified based on itselemental composition. By elemental composition is meant the differenttypes of atoms present in a compound. Thus, the turmeric oil extract ofthe invention can be identified by the amount and/or ratio of thedifferent atoms present in the extract. Methods of determining elementcompositions are well known to the skilled artisan and many commercialentities provide services to determine the elemental compositions. Insome embodiments, the turmeric oil extract comprises from about 70 toabout 75% carbon and from about 5 to about 10% hydrogen. In some furtherembodiments of this, the turmeric oil extract comprises about 73% carbonand 8% hydrogen. In some embodiments, the turmeric oil extract comprisesabout 73.8% carbon and about 8.7% hydrogen.

Alternatively, or in addition, the turmeric oil extract of the inventioncan be identified based on NMR spectra. Accordingly, in someembodiments, at least one compound in the turmeric oil extract has anNMR spectra as shown in FIG. 2, 3, 32-38 or 44. In some embodiments, theturmeric oil extract comprises at least two compounds having an NMRspectra shown in FIG. 2, 3, 32-38, or 44. In one embodiment, theturmeric oil extract comprises a compound having an NMR spectra shown inFIG. 32 and a compound having an NMR spectra shown in FIG. 33. In oneembodiment, the turmeric oil extract comprises a compound having an NMRspectra shown in FIG. 44A and a compound having an NMR spectra shown inFIG. 44B. Methods of predicting NMR spectra for a compound of knownstructure are known to the skilled artisan. For example, commerciallyavailable computer program ACD/NMR Predictors from Advanced ChemistryDevelopment, Inc. (Toronto, Ontario, Canada) can predict the followingnuclei—¹H, ¹³C, ¹⁵N, ¹⁹F, and ³¹P—for 1D spectra, and ¹H and ¹³C (and¹⁵N) for 2D spectrum.

The turmeric oil extract of the invention can also be identified basedon GC mass spectra of the extract. Accordingly, in some embodiments, atleast one compound in the turmeric oil extract has an GC mass spectrumas shown in FIGS. 41-43 and 45-48. It is to be understood, that in someembodiments, at least one compound in the turmeric oil extract has afragmentation pattern as shown in FIG. 41-43 or 45-48. In someembodiments, the turmeric oil extract has an R_(f) of 0.42 by TLC (ethylacetate/hexane 15/85).

The turmeric oil extracts described herein have anti-inflammatoryactivity and/or anti-cancer activity. Furthermore, the turmeric oilextracts are also analgesic. Moreover, as discussed herein, the turmericoil extracts show synergistic anti-inflammatory activity and/oranalgesic activity with anti-inflammatory agents. The turmeric oilextracts described herein also enhance the anti-cancer activity ofanti-cancer agents and show a synergistic effect with anti-canceragents. In some embodiments, the turmeric oil fraction has anti-canceractivity pancreatic, breast or prostate cancer.

In some embodiments, the turmeric oil fraction comprises a mixture ofβ-turmerone/curlone, α-turmerone, and ar-turmerone.

In some embodiments, the turmeric oil fraction comprises at least 35%β-turmerone/curlone. In some embodiments, the turmeric oil fractioncomprises from about 40% to about 65% (e.g., about 40, about 41%, about42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%,about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about55%, about 56%, about 57%, about 58%, about 59%, or about 60%β-turmerone/curlone.

In some embodiments, the turmeric oil fraction comprises at least 20%α-turmerone. In some embodiments, the turmeric oil fraction comprisesfrom about 25% to about 50% (e.g., about 25%, about 26%, about 27%,about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%,about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about47%, about 48%, about 49%, or about 50%) α-turmerone.

In some embodiments, the turmeric oil fraction comprises at least 1%ar-turmerone. In some embodiments, the turmeric oil fraction comprisesfrom about 1% to about 15% (e.g., about 1%, about 2%, about 3%, about4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about11%, about 12%, about 13%, about 14%, or about 15%) ar-turmerone.

In some embodiments, the turmeric oil fraction comprises from about 40%to about 65% β-turmerone/curlone, from about 25% to about 50%α-turmerone, and from about 1% to about 15% ar-turmerone.

In one embodiment, the turmeric oil fraction comprises about 53%β-turmerone/curlone, about 36% α-turmerone, and about 4% ar-turmerone.

In some embodiments, the turmeric oil fraction comprises a mixture ofβ-sesquiphellandrene, 7-epi-zingiberene, and ar-curcumene.

In some embodiments, the turmeric oil fraction comprises at least 35%β-sesquiphellandrene. In some embodiments, the turmeric oil fractioncomprises from about 40% to about 60% (e.g., about 40, about 41%, about42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%,about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about55%, about 56%, about 57%, about 58%, about 59%, or about 60%)β-sesquiphellandrene.

In some embodiments, the turmeric oil fraction comprises at least 15%7-epi-zingiberene. In some embodiments, the turmeric oil fractioncomprises from about 20% to about 40% (e.g., 21%, about 22%, about 23%,about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%,about 37%, about 38%, about 39%, or about 40%) 7-epi-zingiberene.

In some embodiments, the turmeric oil fraction comprises at least 5%ar-curcumene. In some embodiments, the turmeric oil fraction comprisesfrom about 10% to about 30% (e.g. 10%, about 11%, about 12%, about 13%,about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%,about 27%, about 28%, about 29%, about or about 30%) ar-curcumene.

In some embodiments, the turmeric oil fraction comprises from about 40%to about 60% β-sesquiphellandrene, from about 20% to about 40%7-epi-zingiberene, and from about 10% to about 30% ar-curcumene.

In one embodiment, the turmeric oil fraction comprises about 50%β-sesquiphellandrene, about 31% 7-epi-zingiberene, and about 19%ar-curcumene.

In some embodiments, the turmeric oil fraction comprises from about 40%to about 65% ar-turmerone, from about 25% to about 50% α-turmerone, andfrom about 5% to about 20% β-turmerone.

In one embodiment, the turmeric oil fraction comprises about 53%ar-turmerone/curlone, about 36% α-turmerone, and about 9% β-turmerone.

In some embodiments, the turmeric oil fraction comprises from about 55%to about 80% α-turmerone, from about 10% to about 30% β-turmerone, andfrom about 0.5% to about 5% ar-turmerone.

In one embodiment, the turmeric oil fraction comprises about 78%α-turmerone, about 20% β-turmerone, and about 2% ar-turmerone.

It is to be understood that amounts of the various components in theturmeric oil fractions can be based on moles or weights.

Curcumin Derivatives

In another aspect, the invention provides a curcumin derivative havingthe structure shown in formula (I):

wherein: R¹ and R² are independently H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl, apeptide, —C(O)R³, —C(O)OR³, or —C(O)NR³R³, provided that at least one ofR¹ and R² is not H, or provided that both of R¹ and R² are not octyl orhexadecyl, or provided that one of R¹ or R² is not H and the other isnot octyl or hexadecyl; R³ is independently for each occurrence H,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted cyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, optionallysubstituted heteroaryl; and analogs, derivatives, isomers, prodrugs, andpharmaceutically acceptable salts thereof.

One of skill in the art is well aware that curcumin can exist in atleast two tautomeric forms, keto and enol. Without wishing to be boundby theory, the enol form is more energetically stable in the solid phaseand in solution. Accordingly, while curcumin derivatives of formula (I)are shown in the keto form herein, the curcumin derivatives of formula(I) can exist in either the keto or the enol tautomer.

In some embodiment, R¹ and R² are the same.

In some embodiments, one of R¹ and R² is H.

In some embodiments, at least one of R¹ and R² is selected from thegroup consisting of acetyl, myristoleoyl, palmitoleoyl, sapienoyl,oleoyl, linoleoyl, α-linoleoyl, α-linolenoyl, γ-linolenoyl,arcchidionoyl, eicosapentaenoyl, erucoyl, docosahexaenoyl, lauroyl,myrsitoyl, palmitoyl, stearoyl, arachidoyl, behenoyl, lignoceroyl,certoyl and any combinations thereof.

In some embodiments, at least one of R¹ and R² is —C(O)R³, —C(O)OR³, or—C(O)NR³R³.

In some embodiments, R³ is an optionally substituted aryl. Theoptionally substituted aryl can be substituted at the 2-, 3-, 4-, or5-position or any combinations of these positions. One preferredoptionally substituted aryl is a 2-substituted phenyl.

In some embodiments, at least one of R¹ and R² is —C(O)R³ and R³ is anoptionally substituted aryl. Accordingly, at least one of R¹ and R² canbe

wherein R⁴ is H, optionally substituted alkyl, optionally substitutedalkenyl, optionally substituted alkynyl, or optionally substituted acyl.In some embodiments of this, R⁴ is H or acetyl, i.e. R¹ and/or R² can be

In some embodiments, at least one of R¹ and R² is —C(O)R³; and R³ is analkenyl comprising 1, 2, 3, 4, 5 or 6 double bonds. When R³ is analkenyl, it can also comprise 1, 2, 3 or 4 triple bonds in addition tothe double bonds.

In some embodiments, at least one of R¹ and R² is —C(O)R³ and R³ is analkynyl comprising 1, 2, 3, 4, 5 or 6 triple bonds.

In some embodiments, at least one of R¹ and R² is -linker-R⁵, wherein R⁵is a carbohydrate, a peptide, and analogs and derivatives thereof.

As used herein, the term “linker” means an organic moiety that connectstwo parts of a compound. Linkers typically comprise a direct bond or anatom such as oxygen or sulfur, a unit such as NH, C(O), C(O)NH, SO, SO₂,SO₂NH or a chain of atoms, such as substituted or unsubstituted alkyl,substituted or unsubstituted alkenyl, substituted or unsubstitutedalkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl,heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl,heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl,heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl,alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl,alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl,alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl,alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl,alkenylheteroarylalkenyl, alkenylheteroarylalkynyl,alkynylheteroarylalkyl, alkynylheteroarylalkenyl,alkynylheteroarylalkynyl, alkylheterocyclylalkyl,alkylheterocyclylalkenyl, alkylhererocyclylalkynyl,alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl,alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl,alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl,alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl,alkynylhereroaryl, where one or more methylenes can be interrupted orterminated by O, S, S(O), SO₂, N(R¹¹)₂, C(O), cleavable linking group,substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl, substituted or unsubstituted heterocyclic; where R¹¹ ishydrogen, acyl, aliphatic or substituted aliphatic.

In some embodiments, the linker is —C(O)(CH₂)_(m)C(O)O—, wherein m is aninteger from 1 to 10. Preferably m is 1, 2, 3, 4, or 5. Accordingly, atleast one of R¹ and R² can be —C(O)(CH₂)₅C(O)OR⁵.

As used herein, the term “carbohydrate” includes, but is not limited to,compounds that contain oxygen, hydrogen and carbon atoms, typically(C.H₂.O)n wherein n=3. Carbohydrates include, but are not limited to,compounds such as monosaccharides, oligosaccharides, polysaccharides,glycoproteins, glycolipids and the like. The hydroxyl and amino groupsof the monosaccharide can be present as free or as protected groups.Preferred protecting groups include acetonide, t-butoxy carbonyl groups,etc. The monosaccharide can be of L or D configuration. A cyclicmonosaccharide may contain a 5 or 6 membered ring in the α or βconfiguration. Exemplary monosaccharides include, but are not limitedto, glucose, glyceraldehydes, erythrose, threose, ribulose, xylulose,ribose, arabinose, deoxyribose, xylose, lyxose, psicose, fructose,sorbose, tagatose, allose, altrose, mannose, gulose, idose, galactose,talose, fucose, fuculose, rhamnose, sedoheptulose, octose, nonose(Neuraminic acid), and the like.

While a monosaccharide can be linked at any carbon, it is preferred thata hexose is linked at the C4 position. Accordingly, in some embodiments,at least one of R¹ and R² is

wherein R⁶ is OH, amino, mono(C₁-C₆alkyl)amino, di(C₁-C₆alkyl)amino,C₁-C₆alkyl, cyclyl, or alkoxy; and R⁷ is a H, alkyl, cyclyl,heterocyclyl, aryl, heteroaryl, carbohydrate, or a peptide. It is to beunderstood that the pyranose moiety can have the α or β configuration atC1.

In some embodiments, R⁶ is OH.

In some embodiments, R⁷ is H.

In some embodiments, the carbohydrate is glucose and R¹ and/or R² can be

The term “peptide” as used herein is intended to be a generic term whichbroadly includes short peptides (typically less than 100 amino acids).“Peptide” as used generically herein also includes modified peptides.Generally, a peptide of the invention comprises two or more amino acids.A peptide can be linked by its N-terminus, C-terminus, and/or an aminoacid side chain. In some embodiments, the peptide is linked by the sidechain of a lysine amino acid present in the peptide.

In some embodiments, R¹ and R² both are not —C(O)CH₃.

In some embodiments, a curcumin derivative of formula (I) is not acurcumin derivative described in U.S. Pat. App. Pub. No. 2007/0060644,content of which is herein incorporated by reference in its entirety.

In some embodiments, a curcumin derivative of formula (I) is not acurcumin derivative described in Majhi, et al., “Binding of curcumin andits long chain derivatives to the activator binding domain of novelprotein kinase C”, Bioorganic & Medicinal chemistry, 2010, 18:1591-1598, content of which is herein incorporated by reference.

Some exemplary curcumin derivatives of formula (I) are shown FIGS. 1Aand 1B and include di(acetylsalicyloyl)-curcuminmonoacetylsalicyloyl-curcumin, diacetyl-curcumin, monoacetyl-curcumin,diaglutaroyl-curcumin, monoglutaroyl-curcumin,di-gluocose-glutaroyl-curcumin, mono-gluocose-glutaroyl-curcumin,monolinoleol-curcumnin, di-linoleoyl-curcumin and peptide-curcuminconjugates.

The curcumin derivatives described herein have anti-inflammatoryactivity, anti-cancer activity, and/or are analgesic. Furthermore, theturmeric oil extracts are also analgesic. Furthermore, the inventor hasdiscovered that the curcumin derivatives also show a synergistic effectwith anti-inflammatory agents and/or anti-cancer agents.

Aspirin Conjugates

Aspirin (acetylsalicylic acid) is not very water soluble, only 0.33 g in100 mL. Accordingly, some of the undesirable side effects of aspirinresult from undissolved particles in the gastrointestinal mucosa causingulcers and bleeding. The inventor has discovered thatcarbohydrate-aspirin conjugates unexpectedly have enhanced anti-cancerand anti-inflammatory activity relative to unconjugated aspirin.Accordingly, in one aspect, the invention provides compounds of formula(II) for treatment of inflammation, an inflammatory disease orcondition, or cancer in subject in need thereof.

A compound of formula (II), also referred to as a carbohydrate-salicylicacid conjugate, a salicylic, a carbohydrate-aspirin conjugate or anaspirin conjugate herein, has the following structure:

wherein: R⁸ is a carbohydrate; R⁹ is H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl, oroptionally substituted acyl; and analogs, derivatives, isomers,prodrugs, and pharmaceutically acceptable salts thereof.

In some embodiments, R⁸ is selected from the group consisting ofglucose, glyceraldehydes, erythrose, threose, ribulose, xylulose,ribose, arabinose, deoxyribose, xylose, lyxose, psicose, fructose,sorbose, tagatose, allose, altrose, mannose, gulose, idose, galactose,talose, fucose, fuculose, rhamnose, sedoheptulose, octose, nonose(Neuraminic acid), and the like.

In some embodiments, R⁹ is H or acetyl.

In some embodiments, a compound of formula (II) is of formula (IIa):

wherein: R¹⁰ is OH, amino, mono(C₁-C₆alkyl)amino, di(C₁-C₆alkyl)amino,C₁-C₆alkyl, cyclyl, or alkoxy; and R¹¹ is H, alkyl, cyclyl,heterocyclyl, aryl, heteroaryl, carbohydrate, or a peptide.

In some embodiments, R¹⁰ is OH.

In some embodiments, R¹¹ is H or an acyl, e.g., C₁-C₆acyl such asacetyl.

In some embodiments, a compound of formula (II) has the structure shownin formula (IIb):

The compounds of formula (II) have enhanced anti-cancer and/oranti-inflammatory activity relative to the anti-cancer and/oranti-inflammatory activity of aspirin. Accordingly, in some embodiments,a compound of formula (II) is at least 10%, at least 20%, at least 30,at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, at least 95%, or at least 100% more active than aunconjugated aspirin under the same conditions. The activity can be asdetermined in vitro or in vivo.

Furthermore, the compounds of formula (II) have increased solubilityrelative to the solubility of unconjugated aspirin. Accordingly, in someembodiments, a compound of formula (II) is at least 10%, at least 20%,at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, at least 95%, or at least 100% more solublethan aspirin under the same conditions. The at least amount can be basedon weight and/or moles.

Compounds of formula (II) are also described in U.S. Pat. Nos.3,279,990; 4,241,055; 4,242,330; 4,975,269; 5,157,030; 5,550,762;5,700,784; and 5,723,453, Int. Pat. App. Pub. No. WO/1986/003206, andFrench Pat. No. FR M1453, content of all of which is herein incorporatedby reference in its entirety.

Compositions Comprising Turmeric Oil or Turmeric Oil Extract

The inventor has also discovered that turmeric oil and/or a turmeric oilextract enhances the anti-inflammatory activity of anti-inflammatoryagents and enhances the anti-cancer activity of anti-cancer agents.Accordingly, in another aspect, the invention provides a compositioncomprising turmeric oil or a turmeric oil extract and a compoundselected from the group consisting of anti-cancer agents, ananti-inflammatory agents, curcumin, curcumin derivatives (e.g. compoundsof formula (I)), curcumin derivatives, salicylic acid conjugates (e.g.compounds of formula (II)), fish oil, fish oil extract, aspirin and anycombinations thereof, wherein the curcumin ether derivative is offormula (III):

-   -   wherein R₁, R₂, R₃, and R₄ are independently H, alky, acyl,        trialkylsilyl (—Si(alkyl)₃), aryl dialkylsilyl        (—Si(alkyl)₂aryl), diarylalkylsilyl (—Si(aryl)₂alkyl), or        triarylsilyl (—Si(aryl)₃), each of which can be optionally        substituted.

In some compounds of formula (III), at least one of (e.g., one, two,three, or four) R₁, R₂, R₃, and R₄ is trialkylsilyl (—Si(alkyl)₃), aryldialkylsilyl (—Si(alkyl)₂aryl), diarylalkylsilyl (—Si(aryl)₂alkyl), ortriarylsilyl (—Si(aryl)₃).

In some compounds of formula (III), each of R₁, R₂, R₃, and R₄ isindependently selected from trialkylsilyl (—Si(alkyl)₃), aryldialkylsilyl (—Si(alkyl)₂aryl), diarylalkylsilyl (—Si(aryl)₂alkyl), ortriarylsilyl (—Si(aryl)₃).

In some compounds of formula (III), each of R₁, R₂, R₃, and R₄ isindependently selected trialkylsilyl (—Si(alkyl)₃).

In some compounds of formula (III), R₂ and R₄ are both alkyl, and R₁ andR₃ are both alkyl or one of R₁ and R₃ is H and the other is alkyl.

In some compounds of formula (III), R₂ and R₄ are both CH₃; and R₁ andR₃ are both alkyl or one of R₁ and R₃ is H and the other is alkyl.

In some compounds of formula (III), at least one of R₁ or R₃ is octyl,hexadecyl, or octadecyl.

In some compounds of formula (III), both of R₁ and R₃ are octyl,hexadecyl, or octadecyl.

In some compounds of formula (III), at least one of (e.g., one, two,three, or four) R₁, R₂, R₃, and R₄ is acyl.

Generally, the turmeric oil or the turmeric oil extract and the othercomponent can be present in any ratio (weight ratio or molar ratio) inthe composition. Accordingly, the turmeric oil or the turmeric oilextract and the other component can be present at a ratio of 99:1 to1:99. In some embodiments, the composition comprises turmeric oil or theturmeric extract and the other compound in a 90:10 to 10:9, 80:20 to20:80, 70:30 to 30:70, 60:40 to 40:60, or 50:50 ratio. In someembodiments, the turmeric oil or turmeric oil extract and the compoundratio is 5:1 to 15:1. In one embodiment, the turmeric oil or turmericoil extract and the compound ratio is 10:1. In one embodiment, ratio ofthe turmeric oil or turmeric oil extract to the compound is 1:1.

In some embodiments, in addition to the turmeric oil and/or turmeric oilextract, the composition comprises two of: anti-cancer agents, ananti-inflammatory agents, curcumin, curcumin derivatives (e.g. compoundsof formula (I)), curcumin ether derivatives (e.g. compounds of formula(III)), salicylic acid conjugates (e.g. compounds of formula (II)), fishoil, fish oil extract, and aspirin.

In some embodiments, in addition to the turmeric oil and/or turmeric oilextract, the composition comprises three of: anti-cancer agents, ananti-inflammatory agents, curcumin, curcumin derivatives (e.g. compoundsof formula (I)), curcumin ether derivatives (e.g. compounds of formula(III)), salicylic acid conjugates (e.g. compounds of formula (II)), fishoil, fish oil extract, and aspirin.

When the composition comprises more than one of the above mentionedcompounds, ratio of each compound to the turmeric oil or the turmericoil can be determined separately or ratio for the total of the compoundscan be determined. Accordingly, in some embodiments, the compositioncomprises at least 50%, at least 60%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, or at least 95% turmeric oiland/or turmeric oil extract.

Some exemplary compositions of this aspect include, but are not limitedto, turmeric oil and aspirin; turmeric oil and a compound of formula(II); turmeric oil and a compound of formula (Ha); turmeric oil andcurcumin; turmeric oil and a compound of formula (I); turmeric oil and acompound of formula (III); turmeric oil anddi(acetylsalicyloyl)-curcumin; turmeric oil andmonoacetylsalicyloyl-curcumin; turmeric oil and diacetyl-curcumin;turmeric oil and monoacetyl-curcumin; turmeric oil anddiaglutaroyl-curcumin; turmeric oil and monoglutaroyl-curcumin; turmericoil and di-gluocose-glutaroyl-curcumin; turmeric oil andmono-gluocose-glutaroyl-curcumin; turmeric oil andmonolinoleol-curcumnin; turmeric oil and di-linoleoyl-curcumin; turmericoil and an anticancer agent; turmeric oil and an anti-inflammatoryagent; turmeric oil and fish oil; turmeric oil and fish oil extract;turmeric oil and a curcumin diglutarate monolipid conjugate; turmericoil and a curcumin diglutarate-dilipid conjugate; turmeric oil and acurcumin-diglutarate-distearin monoester; turmeric oil, a curcumindiglutarate monolipid conjugate and an anti-cancer agent; turmeric oil,a curcumin diglutarate monolipid conjugate and a compound of formula(II); turmeric oil, a curcumin diglutarate monolipid conjugate and ananti-inflammatory agent; turmeric oil, a curcumin diglutarate monolipidconjugate and a compound of formula (II); turmeric oil and aspirin;turmeric oil, aspirin and a curcumin diglutarate monolipid conjugate;turmeric oil and glucose-aspirin conjugate; turmeric oil,aspirin-glucose conjugate and a curcumin diglutarate monolipidconjugate; turmeric oil, an anti-cancer agent and an anti-inflammatoryagent; turmeric oil, an anti-cancer agent and a compound of formula (I);turmeric oil, an anti-cancer agent and a compound of formula (III);turmeric oil, an anti-cancer agent and a compound of formula (II);turmeric oil, an anti-inflammatory agent and a compound of formula (I);turmeric oil, an anti-inflammatory agent and a compound of formula(III); turmeric oil, an anti-inflammatory agent and a compound offormula (II); turmeric oil, a compound of formula (I) and a compound offormula (II); turmeric oil, a compound of formula (III) and a compoundof formula (II); turmeric oil and aspirin; turmeric oil, aspirin andcurcumin; turmeric oil and glucose-aspirin conjugate; and turmeric oil,aspirin-glucose conjugate and curcumin. While these exemplarycompositions recite turmeric oil, it is to be understood that a turmericextract can be substituted for the turmeric oil or added in addition tothe turmeric oil in these exemplary compositions.

In some embodiments, the composition comprises a turmeric oil fractionand paclitaxel or a paclitaxel-carbohydrate conjugate. In some furtherembodiments of this, the composition comprises a turmeric oil fractionand a paclitaxel-carbohydrate conjugate selected from the groupconsisting of 2′-(GABA-succinoyl)paclitaxel,2′-(glucose-GABA-succinoyl)paclitaxel, 2′-(glucose-succinoyl)paclitaxel,2′-(glucose-glutamyl)paclitaxel,2′-(glucosamide-GABA-succinoyl)paclitaxel,2′-(glucosamide-succinoyl)paclitaxel,2′-(glucosamide-glutamyl)paclitaxel, 7-(GABA-succinoyl)paclitaxel,7-(glucose-GABA-succinoyl)paclitaxel, 7-(glucose-succinoyl)paclitaxel,7-(glucose-glutamyl)paclitaxel,7-(glucosamide-GABA-succinoyl)paclitaxel,7-(glucosamide-succinoyl)paclitaxel, and7-(glucosamide-glutamyl)paclitaxel.

Without wishing to be bound by a theory, combining an anti-inflammatoryor an anti-cancer agent with turmeric oil and/or a turmeric oil extractleads to enhanced activity for the anti-inflammatory or the anti-canceragent. Accordingly, in some embodiments, the activity of ananti-inflammatory or an anti-cancer agent is increased by at least 10%,at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, atleast 70%, at least 80%, at least 90%, at least 1-fold, at least 2-fold,at least 4-fold, at least 5-fold or more, relative to the activity ofthe anti-inflammatory, the anti-cancer agent, the turmeric oil or theturmeric oil extract alone. The skilled artisan is well aware of methodsand assays for measuring anti-inflammatory or anti-cancer activity of acompound. For example, anti-inflammatory activity can be measured usingan assay designed to test the ability of a compound to antagonize thelocal edema which is characteristic of the inflammatory response.Examples of such assays include, but are not limited to, the carrageenanrat edema test, the oxazolone-induced inflamed mouse ear test, and thearachidonic acid-induced inflamed mouse ear test. Similarly, anti-canceractivity can be determined, for example, by using the LIVE/DEAD® CellViability Assays from Invitrogen (Carlsbad, Calif., USA). Withoutwishing to be bound by a theory, the turmeric oil and/or the turmericoil extract has a synergistic anti-inflammatory, analgesic and/oranti-cancer activity in combination with an anti-inflammatory and/oranti-cancer agent.

In some embodiments, the turmeric oil fraction increases activity ofpaclitaxel, e.g., anti-cancer activity, by at least 10%, at least 20%,at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, at least 1-fold, at least 2-fold, at least4-fold, at least 5-fold or more, relative to the activity of paclitaxelalone.

Compositions Comprising Curcumin or Curcumin Derivative

In one aspect, the invention provides a composition comprising curcumin,a curcumin derivative of formula (I), or a curcumin ether derivative offormula (III) and a compound selected from the group consisting ofanti-cancer agents, anti-inflammatory agents, compounds of formula (II),fish oil, fish oil extract, aspirin and any combinations thereof.Generally, curcumin or curcumin derivate and the other component can bepresent in any ratio (weight ratio or molar ratio) in the composition.Accordingly, the curcumin or the curcumin derivative and the othercomponent can be present at a ratio of 99:1 to 1:99. In someembodiments, the composition comprises the curcumin or the curcuminderivative and the other compound in a 90:10 to 10:90, 80:20 to 20:80,70:30 to 30:70, 60:40 to 40:60, or 50:50 ratio. In some embodiments, thecurcumin or the curcumin derivative and the compound ratio is 5:1 to15:1. In one embodiment, the curcumin or the curcumin derivative and thecompound ratio is 10:1. In one embodiment, the curcumin or the curcuminderivative and the compound ratio is 1:1. In some embodiments, thecomposition comprises curcumin and fish oil in a 2:1 ratio by weight.

In some embodiments, the composition comprises curcumin or curcuminderivative, fish oil and lecithin. In some further embodiments of this,the curcumin or curcumin derivative, fish oil and lecithin are presentin a ratio of 1-10:1-10:1-10 by weight. In one embodiment, the curcuminor curcumin derivative, fish oil and lecithin are present in a ratio of4:2.5:1 by weight. Without wishing to be bound by theory, lecithin canincrease the mixing ability of the various components of the compositionand also has its own beneficial effect.

The term “lecithin” is used herein in its art-recognized manner. See,for example, the United States Pharmacopeia/National Formulary,published by the United States Pharmacopeial Convention, Inc.(Rockville, Md.). The term lecithin as conventionally used in the artrefers to pure phosphatidyl choline and also to crude phospholipidmixtures, containing a variety of other compounds such as fatty acids,triglycerides, sterols, carbohydrates and glycolipids.

Lecithin includes a complex mixture of acetone-insoluble phosphatides,of which phosphatidylcholine is a significant component. The termlecithin is also used as a synonym for phosphatidylcholine. Commerciallysupplied lecithin is typically derived from egg yolk, soybeans, or corn.As used herein, the term “lecithin” encompasses phosphatidyl cholineobtained naturally or synthetically, including de-oiled or de-gummedproducts; derivatives of lecithin and combinations of various types oflecithin. Commercial lecithin is currently available in more than fortydifferent formulations (from sources such as American Lecithin Co.;Lucas Meyer Inc. and Central Soya Inc. among others) varying from crudeoily extracts from natural sources to purified and syntheticphospholipids, all intended to be encompassed by the term “lecithin” asused herein.

Lecithin can be made more hydrophilic by hydroxylation of unsaturatedfatty acid constituents, fractionation or compounding with dispersingagents. Moreover, lecithin may be hydroxylated by treating thephosphatides with hydrogen peroxide or peracids in the presence ofwater-soluble aliphatic carboxylic acids. Alternatively, lecithin mayalso be hydrolyzed enzymatically to yield a powdered soybean lecithin.Another lecithin derivative is lysolecithin, which results from theinteraction of the enzyme phospholipase with lecithin, for example inpancreatic juices. Therefore lecithin derivatives are compounds whichcan be the result of hydroxylation or enzymatic reaction, as mentionedabove or other chemical modification of lecithin, included in the broadterm “lecithin”.

Some examples of suitable lecithins available from Central Soya Inc. ofIowa include BLENDMAX, CENTROBAKE, CENTROCAP, CENTROL CA, CENTROLENE,CENTROMIX, CENTROPHASE, CENTROPHIL, NATHIN and PRECEPT. These names mayrepresent either a single lecithin, or a series of lecithin products,all of which are considered useful for the purposes described herein.

When the composition comprises more than one of the above mentionedcompounds, ratio of each compound to the curcumin or curcumin derivativecan be determined separately or ratio for the total of the compounds canbe determined. Accordingly, in some embodiments, the compositioncomprises at least 50%, at least 60%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, or at least 95% curcuminderivative.

Without wishing to be bound by theory, a composition described hereinmay comprise a small amount (0.1%-5%) of curcumin in the composition asan antioxidant to preserve the composition.

Some exemplary compositions of this aspect include, but are not limitedto, curcumin derivative and aspirin; curcumin derivative and a compoundof formula (II); curcumin derivative and curcumin; curcumin etherderivative and aspirin; curcumin ether derivative and a compound offormula (II); aspirin and di(acetylsalicyloyl)-curcumin; aspirin andmonoacetylsalicyloyl-curcumin; aspirin and diacetyl-curcumin; aspirinand monoacetyl-curcumin; aspirin and diaglutaroyl-curcumin; aspirin andmonoglutaroyl-curcumin; aspirin and di-gluocose-glutaroyl-curcumin;aspirin and mono-gluocose-glutaroyl-curcumin; aspirin andmonolinoleol-curcumnin; aspirin and di-linoleoyl-curcumin; a compound offormula (II) and di(acetylsalicyloyl)-curcumin; a compound of formula(II) and monoacetylsalicyloyl-curcumin; a compound of formula (II) anddiacetyl-curcumin; a compound of formula (II) and monoacetyl-curcumin; acompound of formula (II) and diaglutaroyl-curcumin; a compound offormula (II) and monoglutaroyl-curcumin; a compound of formula (II) anddi-gluocose-glutaroyl-curcumin; a compound of formula (II) andmono-gluocose-glutaroyl-curcumin; a compound of formula (II) andmonolinoleol-curcumnin; a compound of formula (II) anddi-linoleoyl-curcumin; a compound of formula (IIb) anddi(acetylsalicyloyl)-curcumin; a compound of formula (IIb) andmonoacetylsalicyloyl-curcumin; a compound of formula (IIb) anddiacetyl-curcumin; a compound of formula (IIb) and monoacetyl-curcumin;a compound of formula (IIb) and diaglutaroyl-curcumin; a compound offormula (IIb) and monoglutaroyl-curcumin; a compound of formula (IIb)and di-gluocose-glutaroyl-curcumin; a compound of formula (IIb) andmono-gluocose-glutaroyl-curcumin; a compound of formula (IIb) andmonolinoleol-curcumnin; a compound of formula (IIb) anddi-linoleoyl-curcumin; a compound of formula (IIc) anddi(acetylsalicyloyl)-curcumin; a compound of formula (IIc) andmonoacetylsalicyloyl-curcumin; a compound of formula (IIc) anddiacetyl-curcumin; a compound of formula (IIc) and monoacetyl-curcumin;a compound of formula (IIc) and diaglutaroyl-curcumin; a compound offormula (IIc) and monoglutaroyl-curcumin; a compound of formula (IIc)and di-gluocose-glutaroyl-curcumin; a compound of formula (IIc) andmono-gluocose-glutaroyl-curcumin; a compound of formula (IIc) andmonolinoleol-curcumnin; a compound of formula (IIc) anddi-linoleoyl-curcumin; curcumin and aspirin; curcumin andglucose-aspirin conjugate; curcumin and fish oil; and curcumin, fish oiland lecithin;

In some embodiments, the composition comprises a curcumin derivative offormula (I), or a curcumin ether derivative of formula (III) andpaclitaxel or a paclitaxel-carbohydrate conjugate. In some furtherembodiments of this, the composition comprises a curcumin derivative offormula (I), or a curcumin ether derivative of formula (III) and apaclitaxel-carbohydrate conjugate selected from the group consisting of2′-(GABA-succinoyl)paclitaxel, 2′-(glucose-GABA-succinoyl)paclitaxel,2′-(glucose-succinoyl)paclitaxel, 2′-(glucose-glutamyl)paclitaxel,2′-(glucosamide-GABA-succinoyl)paclitaxel,2′-(glucosamide-succinoyl)paclitaxel,2′-(glucosamide-glutamyl)paclitaxel, 7-(GABA-succinoyl)paclitaxel,7-(glucose-GABA-succinoyl)paclitaxel, 7-(glucose-succinoyl)paclitaxel,7-(glucose-glutamyl)paclitaxel,7-(glucosamide-GABA-succinoyl)paclitaxel,7-(glucosamide-succinoyl)paclitaxel, and7-(glucosamide-glutamyl)paclitaxel.

Without wishing to be bound by theory, combining an anti-inflammatory oran anti-cancer agent with curcumin and/or the curcumin derivative leadsto enhanced activity for the anti-inflammatory or the anti-cancer agent.Accordingly, in some embodiments, the activity of an anti-inflammatoryor an anti-cancer agent is increased by at least 10%, at least 20%, atleast 30%, at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, at least 1-fold, at least 2-fold, at least4-fold, at least 5-fold or more, relative to the activity of theanti-inflammatory, the anti-cancer agent, the curcumin or the curcuminderivative alone. Without wishing to be bound by a theory, the curcuminand the curcumin derivative have a synergistic anti-inflammatory,analgesic and/or anti-cancer activity in combination with ananti-inflammatory and/or anti-cancer agent.

In some embodiments, the composition comprises two or more of turmericoil fractions, curcumin, curcumin derivarives, and fish oil. Withoutwishing to be bound by a theory, a composition comprising two or more ofturmeric oil fractions, curcumin, curcumin derivarives, and fish oilhave enhanced anti-inflammatory and analgesic activity relative to theturmeric oil fraction, curcumin, curcumin derivative or fish oil alone.In other words, a composition comprising two or more of turmeric oilfractions, curcumin, curcumin derivarives, and fish oil has asynergistic anti-inflammatory and analgesic activity.

Methods of Use

The extracts, compounds and compositions described herein haveanti-inflammatory, analgesic and/or anti-cancer properties. Accordingly,in one aspect the inventions provides a method of treating a subject forinflammation, or a disease or condition associated with inflammation,the method comprising administering a therapeutically effective amountof a turmeric oil extract, a curcumin derivative, a curcumin etherderivative, a salicylic acid conjugate, turmeric oil extract comprisingcomposition, a curcumin derivative comprising composition, or anycombinations thereof to a subject in need thereof.

As used herein, an anti-inflammation treatment aims to prevent or slowdown (lessen) an undesired physiological change or disorder, such as thedevelopment or progression of the inflammation. Beneficial or desiredclinical results include, but are not limited to, alleviation of asymptom or symptoms, diminishment of extent of disease, stabilized(i.e., not worsening) state of disease, delay or slowing of inflammatorydisease progression, amelioration or palliation of the disease state,and remission (whether partial or total). An anti-inflammatory treatmentcan also mean prolonging survival as compared to expected survival ifnot receiving treatment. In one aspect, an anti-inflammatory treatmentcan completely suppress the inflammatory response.

As used herein, “inflammation” refers to the complex biological responseof vascular tissues to harmful stimuli, such as pathogens, damagedcells, or irritants. Inflammation is a protective attempt by theorganism to remove the injurious stimuli as well as initiate the healingprocess for the tissue. Accordingly, the term “inflammation” includesany cellular process that leads to the production of pro-inflammatorycytokines, inflammation mediators and/or the related downstream cellularevents resulting from the actions of the cytokines thus produced, forexample, fever, fluid accumulation, swelling, abscess formation, andcell death. Pro-inflammatory cytokines and inflammation mediatorsinclude, but are not limited to, IL-1-alpha, IL-1-beta, IL-6, IL-8,IL-11, IL-12, IL-17, IL-18, TNF-alpha, leukocyte inhibitory factor(LIF), IFN-gamma, Oncostatin M (OSM), ciliary neurotrophic factor(CNTF), TGF-beta, granulocyte-macrophage colony stimulating factor(GM-CSF), and chemokines that chemoattract inflammatory cells. As usedherein, “inflammation” also refers to any cellular process that leads tothe activation of caspase-1 or caspase-5.

As used herein, the term “inflammation” refers to both acute responses(i.e., responses in which the inflammatory processes are active) andchronic responses (i.e., responses marked by slow progression andformation of new connective tissue). Acute and chronic inflammation maybe distinguished by the cell types involved. Acute inflammation ofteninvolves polymorphonuclear neutrophils; whereas chronic inflammation isnormally characterized by a lymphohistiocytic and/or granulomatousresponse.

As used herein, the term “inflammation” includes reactions of both thespecific and non-specific defense systems. A specific defense systemreaction is a specific immune system reaction response to an antigen(possibly including an autoantigen). A non-specific defense systemreaction is an inflammatory response mediated by leukocytes incapable ofimmunological memory. Such cells include granulocytes, macrophages,neutrophils and eosinophils. Examples of specific types of inflammationare diffuse inflammation, focal inflammation, croupous inflammation,interstitial inflammation, obliterative inflammation, parenchymatousinflammation, reactive inflammation, specific inflammation, toxicinflammation and traumatic inflammation.

As used herein, the term “specific defense system” is intended to referto that component of the immune system that reacts to the presence ofspecific antigens. Inflammation is said to result from a response of thespecific defense system if the inflammation is caused by, mediated by,or associated with a reaction of the specific defense system. Examplesof inflammation resulting from a response of the specific defense systeminclude the response to antigens such as rubella virus, autoimmunediseases such as lupus erythematosus, rheumatoid arthritis, Reynaud'ssyndrome, multiple sclerosis etc., delayed type hypersensitivityresponse mediated by T-cells, etc. Chronic inflammatory diseases and therejection of transplanted tissue and organs are further examples ofinflammatory reactions of the specific defense system.

As used herein, a reaction of the “non-specific defense system” isintended to refer to a reaction mediated by leukocytes incapable ofimmunological memory. Such cells include granulocytes and macrophages.As used herein, inflammation is said to result from a response of thenonspecific defense system, if the inflammation is caused by, mediatedby, or associated with a reaction of the non-specific defense system.Examples of inflammation which result, at least in part, from a reactionof the non-specific defense system include inflammation associated withconditions such as: adult respiratory distress syndrome (ARDS) ormultiple organ injury syndromes secondary to septicemia or trauma;reperfusion injury of myocardial or other tissues; acuteglomerulonephritis; reactive arthritis; dermatoses with acuteinflammatory components; acute purulent meningitis or other centralnervous system inflammatory disorders; thermal injury; hemodialysis;leukophoresis; ulcerative colitis; Crohn's disease; necrotizingenterocolitis; granulocyte transfusion associated syndromes; andcytokine-induced toxicity. The term immune-mediated refers to a processthat is either autoimmune or inflammatory in nature.

The term “inflammatory diseases” refers to diseases and conditionsassociated with inflammation. Exemplary inflammatory diseases include,but are not limited to, rheumatoid arthritis, inflammatory boweldisease, pelvic inflammatory disease, ulcerative colitis, psoriasis,systemic lupus erythematosus, multiple sclerosis, type 1 diabetesmellitus, psoriasis, vaculitis, allergic inflammation such as allergicasthma, atopic dermatitis, and contact hypersensitivity. Other examplesof inflammatory diseases or disorders include, but are not limited to,rheumatoid arthritis, multiple sclerosis (MS), systemic lupuserythematosus, Graves' disease (overactive thyroid), Hashimoto'sthyroiditis (underactive thyroid), Type 1 diabetes mellitus, celiacdisease, Crohn's disease and ulcerative colitis, Guillain-Barresyndrome, primary biliary sclerosis/cirrhosis, sclerosing cholangitis,autoimmune hepatitis, Raynaud's phenomenon, scleroderma, Sjogren'ssyndrome, Goodpasture's syndrome, Wegener's granulomatosis, polymyalgiarheumatica, temporal arteritis/giant cell arteritis, chronic fatiguesyndrome CFS), psoriasis, autoimmune Addison's Disease, ankylosingspondylitis, Acute disseminated encephalomyelitis, antiphospholipidantibody syndrome, aplastic anemia, idiopathic thrombocytopenic purpura,Myasthenia gravis, opsoclonus myoclonus syndrome, optic neuritis, Ord'sthyroiditis, pemphigus, pernicious anaemia, polyarthritis in dogs,Reiter's syndrome, Takayasu's arteritis, warm autoimmune hemolyticanemia, Wegener's granulomatosis, fibromyalgia (FM), autoinflammatoryPAPA syndrome, Familial Mediterranean Fever, familial coldautoinflammatory syndrome, Muckle-Wells syndrome, the neonatal onsetmultisystem inflammatory disease, inflammatory or allergic diseases(e.g., systemic anaphylaxis or hypersensitivity responses, drugallergies, insect sting allergies), inflammatory bowel diseases (e.g.,colitis, ileitis and enteritis; vaginitis), psoriasis and inflammatorydermatoses (e.g., dermatitis, eczema, atopic dermatitis, allergiccontact dermatitis, urticaria, vasculitis, spondyloarthropathies,scleroderma, respiratory allergic diseases (e.g., asthma, allergicrhinitis, hypersensitivity lung diseases), autoimmune diseases (e.g.arthritis (rheumatoid and psoriatic), osteoarthritis, multiplesclerosis, systemic lupus erythematosus, diabetes mellitus,glomerulonephritis, and the like), graft rejection (including allograftrejection and graft-v-host disease), and other diseases in whichundesired inflammatory responses are to be inhibited (e.g., myositis,inflammatory CNS disorders such as stroke and closed-head injuries,neurodegenerative diseases, Alzheimer's disease, encephalitis,meningitis, osteoporosis, gout, hepatitis, nephritis, sepsis,sarcoidosis, conjunctivitis, otitis, chronic obstructive pulmonarydisease, sinusitis and Bechet's syndrome), chronic prostatitis,glomerulonephritis, inflammatory bowl diseases, pelvic inflammatorydisease, reperfusion injury, silicosis, vasculitis, inflammatorymyopathies, hypersensitivities, migraine, psoriasis, gout, andatherosclerosis.

Without wishing to be bound by theory, one mechanism by whichadministration of turmeric oil extract and/or curcumin derivatives maytreat disease is through inhibition of the activity of AP-1, NF-κBand/or GSTP1-1 Inhibition of NF-κB results in a decrease in NF-κBactivity, and includes direct inhibition and indirect inhibition. Directinhibition is the direct effect of a turmeric oil extract and/or acurcumin derivative on NF-κB and its activity. For example, one type ofdirect inhibition of NF-κB is a block of NF-κB DNA interactions.Indirect inhibition, on the other hand, involves the effect of aturmeric oil extract and/or a curcumin derivative on other compoundsinvolved in the regulation of NF-κB that leads to a decrease in NF-κBactivity. For example, as phosphorylation of the NF-κB regulator IκB byIκB kinases (IKK) or Src family kinases (SFK) results in a dysregulationof NF-κB, and an accompanying increase in NF-κB activity, inhibition ofIKK or SFK by turmeric oil extracts and/or curcumin derivatives providesan example of indirect inhibition.

Inhibition of AP-1 results in a decrease in AP-1 activity, and includesdirect inhibition and indirect inhibition. Direct inhibition is thedirect effect of a turmeric oil extract and/or a curcumin derivative onAP-1 (or its subunits) and its activity. Indirect inhibition, on theother hand, involves the effect of a turmeric oil extract and/or acurcumin derivative on other compounds involved in the regulation ofAP-1 that leads to a decrease in AP-1 activity. For example, indirectinhibition of AP-1 activity may occur as a result of an affect on AP-1activating proteins such as mitogen-activated protein kinases (MAPK) orc-Fos-regulating kinase (FRK).

Inhibition of GSTP1-1 results in a decrease GSTP1-1 activity, andincludes direct inhibition and indirect inhibition. Direct inhibition isthe direct effect of a turmeric oil extract and/or a curcumin derivativeon GSTP1-1 (or its subunits) and its activity. Indirect inhibition, onthe other hand, involves the effect of a turmeric oil extract and/or acurcumin derivative on other compounds involved in the regulation ofGSTP1-1 that leads to a decrease in GSTP1-1 activity.

Also described herein is a method of treating a subject afflicted withcancer, a precancerous condition and/or metastasis, the methodcomprising administering a therapeutically effective amount of aturmeric oil extract, a curcumin derivative, a curcumin etherderivative, a salicylic acid conjugate, a composition described herein,or any combinations thereof to a subject in need thereof. By “reduced”in the context of cancer is meant reduction of at least 10% in thegrowth rate of a tumor or the size of a tumor or cancer cell burden.

As used herein, an anti-cancer treatment aims to reduce, prevent oreliminate cancer cells or the spread of cancer cells or the symptoms ofcancer in the local, regional or systemic circulation. Anti-cancertreatment also means the direct treatment of tumours, for example byreducing or stabilizing their number or their size (curative effect),but also by preventing the in situ progression of tumour cells or theirdiffusion, or the establishment of tumours; this also includes thetreatment of deleterious effects linked to the presence of such tumours,in particular the attenuation of symptoms observed in a patient or animprovement in quality of life.

As used herein, the term “cancer” refers to an uncontrolled growth ofcells that may interfere with the normal functioning of the bodilyorgans and systems. Cancers that migrate from their original locationand seed vital organs can eventually lead to the death of the subjectthrough the functional deterioration of the affected organs. Ametastasis is a cancer cell or group of cancer cells, distinct from theprimary tumor location resulting from the dissemination of cancer cellsfrom the primary tumor to other parts of the body. At the time ofdiagnosis of the primary tumor mass, the subject may be monitored forthe presence of in transit metastases, e.g., cancer cells in the processof dissemination. As used herein, the term cancer, includes, but is notlimited to the following types of cancer, breast cancer, biliary tractcancer, bladder cancer, brain cancer including glioblastomas andmedulloblastomas; cervical cancer; choriocarcinoma; colon cancer;endometrial cancer; esophageal cancer, gastric cancer; hematologicalneoplasms including acute lymphocytic and myelogenous leukemia; T-cellacute lymphoblastic leukemia/lymphoma; hairy cell leukemia; chromicmyelogenous leukemia, multiple myeloma; AIDS-associated leukemias andadult T-cell leukemia lymphoma; intraepithelial neoplasms includingBowen's disease and Paget's disease; liver cancer; lung cancer;lymphomas including Hodgkin's disease and lymphocytic lymphomas;neuroblastomas; oral cancer including squamous cell carcinoma; ovariancancer including those arising from epithelial cells, stromal cells,germ cells and mesenchymal cells; pancreatic cancer; prostate cancer;rectal cancer; sarcomas including leiomyosarcoma, rhabdomyosarcoma,liposarcoma, fibrosarcoma, and osteosarcoma; skin cancer includingmelanoma, Merkel cell carcinoma, Kaposi's sarcoma, basal cell carcinoma,and squamous cell cancer; testicular cancer including germinal tumorssuch as seminoma, non-seminoma (teratomas, choriocarcinomas), stromaltumors, and germ cell tumors; thyroid cancer including thyroidadenocarcinoma and medullar carcinoma; and renal cancer includingadenocarcinoma, Wilms tumor. Examples of cancer include but are notlimited to, carcinoma, including adenocarcinoma, lymphoma, blastoma,melanoma, sarcoma, and leukemia. More particular examples of suchcancers include squamous cell cancer, small-cell lung cancer, non-smallcell lung cancer, gastrointestinal cancer, Hodgkin's and non Hodgkin'slymphoma, pancreatic cancer, glioblastoma, cervical cancer, ovariancancer, liver cancer such as hepatic carcinoma and hepatoma, bladdercancer, breast cancer, colon cancer, colorectal cancer, endometrialcarcinoma, salivary gland carcinoma, kidney cancer such as renal cellcarcinoma and Wilms' tumors, basal cell carcinoma, melanoma, prostatecancer, vulval cancer, thyroid cancer, testicular cancer, esophagealcancer, and various types of head and neck cancer. Other cancers will beknown to the artisan. In some embodiments, cancer is pancreatic, breastor prostate cancer.

As used herein, the term “precancerous condition” has its ordinarymeaning, i.e., an unregulated growth without metastasis, and includesvarious forms of hyperplasia and benign hypertrophy. Accordingly, a“precancerous condition” is a disease, syndrome, or finding that, ifleft untreated, can lead to cancer. It is a generalized state associatedwith a significantly increased risk of cancer. Premalignant lesion is amorphologically altered tissue in which cancer is more likely to occurthan its apparently normal counterpart. Examples of pre-malignantconditions include, but are not limited to, oral leukoplakia, actinickeratosis (solar keratosis), Barrett's esophagus, atrophic gastritis,benign hyperplasia of the prostate, precancerous polyps of the colon orrectum, gastric epithelial dysplasia, adenomatous dysplasia, hereditarynonpolyposis colon cancer syndrome (HNPCC), Barrett's esophagus, bladderdysplasia, precancerous cervical conditions, and cervical dysplasia.

Without wishing to be bound by theory, administration of a turmeric oilextract, a curcumin derivative, and/or a salicylic acid conjugate caninhibit the activity Glutathione S-transferase P1-1 (GSTP1-1), NFκBand/or AP-1 Inhibition of GSTP1-1 may occur by affecting genetranscription and/or by direct effects on enzyme activity.

Administration of the turmeric oil extract, the curcumin derivative,and/or the salicylic acid conjugate is especially advantageous in caseswhere the cancer cells may develop or have developed resistance to oneor more anti-cancer agents; and/or when the cancer cells overexpressGSTP1-1. Expression of GSTP1-1 may allow the cancer cell to pump out theanti-cancer agent, and by inhibiting the activity of GSTP1-1, theturmeric oil extract, the curcumin derivative, and/or the salicylic acidconjugate can preserve or prolong the cytostatic or cytotoxic effects ofthe anti-cancer agent. Compositions described herein are particularlyuseful for improving the effectiveness of anti-cancer agents bypreventing GSTP1-1's inhibition of pro-apoptotic factors, particularlyc-Jun N-terminal kinase (JNK).

Curcumin was shown recently to inhibit apoptosis in cancer cells in partthrough its ability to inhibit the expression of GSTP1-1 mRNA andprotein, which was demonstrated to be the result of inhibition of theactivation of NFκB. This observation that compounds such as curcumin canblock activation of NFκB raises the possibility that synthetic drugs canbe developed that are more potent than curcumin, and that these drugswill promote apoptosis in cancer cells. These drugs could sensitizecancer cells to conventional adjuvant chemotherapy by blocking theNFκB-dependent development of the anti-apoptotic prosurvival state, andinhibit the expression of GSTP1-1. In addition, curcumin inhibits theGSTP1-1 catalyzed conjugation of glutathione with electrophiles.Furthermore, curcumin inhibits the proliferation of a variety of tumorcells and has anti-metastatic activity, possibly owing to its ability toinduce apoptosis by inhibiting NFKB.

Curcumin contains two alpha, beta-unsaturated carbonyl groups, one ofwhich exists as the enol tautomer. Curcumin reacts with glutathione;this reaction is accelerated by GSTP1-1, indicating that curcumin is asubstrate of GSTP1-1, albeit a poor substrate. Curcumin also inhibitsGSTP1-1 in its conjugation of glutathione with other electrophiles,suggesting that curcumin is both a substrate and an inhibitor ofGSTP1-1. This is consistent with the known inhibition of GSTP1-1 by theflavonoid quercetin, which, like curcumin, is also a polyphenol.Curcumin itself has low bioavailability and therefore is not a promisingdrug.

Derivatives of curcumin with good bioavailability can provide newanti-cancer drugs that, for example, can inhibit the catalytic activityof GSTP1-1, thereby sensitizing cancer cells to conventionalchemotherapy by drugs that normally are metabolized through GSTP1-1catalyzed conjugation with glutathione; and/or, through inhibition ofGSTP1-1 and/or NFκB the curcumin derivatives, contribute to improvedchemotherapeutic drug sensitivity of cancer cells by promoting thepro-apoptotic state. While not wishing to be bound by theory, thesederivatives thus may have a dual mechanism of action—both the inhibitionof the catalytic activity of GSTP1-1 and the down-regulation of GSTP1-1transcription through inhibition of NFκB. GSTP1-1 inhibitors may limitthe ability of GSTP1-1 to inactivate other cancer drugs and can prove tobe synergistic when combined with other anti-cancer agents.

As used herein, a “subject” means a human or animal. Examples ofsubjects include primates (e.g., humans, and monkeys). Usually theanimal is a vertebrate such as a primate, rodent, domestic animal orgame animal. Primates include chimpanzees, cynomologous monkeys, spidermonkeys, and macaques, e.g., Rhesus. Rodents include mice, rats,woodchucks, ferrets, rabbits and hamsters. Domestic and game animalsinclude cows, horses, pigs, deer, bison, buffalo, feline species, e.g.,domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g.,chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon.Patient or subject includes any subset of the foregoing, e.g., all ofthe above, but excluding one or more groups or species such as humans,primates or rodents. In certain embodiments of the aspects describedherein, the subject is a mammal, e.g., a primate, e.g., a human. Theterms, “patient” and “subject” are used interchangeably herein. Theterms, “patient” and “subject” are used interchangeably herein. Asubject can be male or female. A subject can be one who has not beenpreviously diagnosed with inflammation, inflammatory disease orcondition, and/or cancer.

Preferably, the subject is a mammal. The mammal can be a human,non-human primate, mouse, rat, dog, cat, horse, or cow, but are notlimited to these examples. Mammals other than humans can beadvantageously used as subjects that represent animal models ofinflammatory disease or disorder, or as animal models of cancer. Inaddition, the methods and compositions described herein can be used totreat domesticated animals and/or pets.

A subject can be one who has been previously diagnosed withinflammation, inflammatory disease or condition, and/or cancer. Withoutwishing to be bound by a theory, a subject can be diagnosed as having aninflammatory disease by having increased levels of at least oneinflammatory disease marker compared to the level of that marker in asubject not having the inflammatory disease. Pro-inflammatory cytokinesand inflammation mediators include, but are not limited to, IL-1-alpha,IL-1-beta, IL-6, IL-8, IL-11, IL-12, IL-17, IL-18, TNF-alpha, leukocyteinhibitory factor (LIF), IFN-gamma, Oncostatin M (OSM), ciliaryneurotrophic factor (CNTF), TGF-beta, granulocyte-macrophage colonystimulating factor (GM-CSF), and chemokines that chemoattractinflammatory cells. A number of assays for in vivo state of inflammationare known in the art. See for example U.S. Pat. Nos. 5,108,899 and5,550,139, contents of both of which are herein incorporated byreference.

A method described herein can further comprise selecting a subject whohas inflammation, inflammatory disease or condition, and/or cancer. Themethod can also comprise the step of diagnosing a subject forinflammation, inflammatory disease or condition, and/or cancer beforeonset of administration or treatment regime.

The phrase “therapeutically-effective amount” as used herein means thatamount of a compound, material, or composition comprising a conjugatewhich is effective for producing some desired therapeutic effect in atleast a sub-population of cells in an animal at a reasonablebenefit/risk ratio applicable to any medical treatment.

Determination of a therapeutically effective amount is well within thecapability of those skilled in the art. Generally, a therapeuticallyeffective amount can vary with the subject's history, age, condition,sex, as well as the severity and type of the medical condition in thesubject, and administration of other pharmaceutically active agents.

As used herein, the term “administer” refers to the placement of acomposition into a subject by a method or route which results in atleast partial localization of the composition at a desired site suchthat desired effect is produced. Routes of administration suitable forthe methods of the invention include both local and systemicadministration. Generally, local administration results in more of thecomposition being delivered to a specific location as compared to theentire body of the subject, whereas, systemic administration results indelivery to essentially the entire body of the subject.

A conjugate described herein can be administered by any appropriateroute known in the art including, but not limited to, oral or parenteralroutes, including intravenous, intramuscular, subcutaneous, transdermal,airway (aerosol), pulmonary, nasal, rectal, vaginal, and topical(including on the skin, and body cavities, such as buccal, vaginal,rectal and sublingual) administration.

Exemplary modes of administration include, but are not limited to,injection, infusion, instillation, inhalation, or ingestion. “Injection”includes, without limitation, intravenous, intramuscular, intraarterial,intrathecal, intraventricular, intracapsular, intraorbital,intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous,subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal,intracerebro spinal, and intrasternal injection and infusion. In someembodiments of the aspects described herein, the compositions areadministered by intravenous infusion or injection. In some embodiments,administration is oral.

Pharmaceutical Compositions

For administration to a subject, a compound and/or composition describedherein can be provided in pharmaceutically acceptable compositions.Accordingly, in one aspect, the invention provides a pharmaceuticalcomposition comprising one or more of a compound and/or compositiondescribed herein, formulated together with one or more pharmaceuticallyacceptable carriers (additives) and/or diluents. As described in detailbelow, the pharmaceutical compositions of the present invention can bespecially formulated for administration in solid or liquid form,including those adapted for the following: (1) oral administration, forexample, drenches (aqueous or non-aqueous solutions or suspensions),gavages, lozenges, dragees, capsules, pills, tablets (e.g., thosetargeted for buccal, sublingual, and systemic absorption), boluses,powders, granules, pastes for application to the tongue; (2) parenteraladministration, for example, by subcutaneous, intramuscular, intravenousor epidural injection as, for example, a sterile solution or suspension,or sustained-release formulation; (3) topical application, for example,as a cream, ointment, or a controlled-release patch or spray applied tothe skin; (4) intravaginally or intrarectally, for example, as apessary, cream or foam; (5) sublingually; (6) ocularly; (7)transdermally; (8) transmucosally; or (9) nasally. Additionally,compounds can be implanted into a patient or injected using a drugdelivery system. See, for example, Urquhart, et al., Ann. Rev.Pharmacol. Toxicol. 24: 199-236 (1984); Lewis, ed. “Controlled Releaseof Pesticides and Pharmaceuticals” (Plenum Press, New York, 1981); U.S.Pat. No. 3,773,919; and U.S. Pat. No. 35 3,270,960, content of all ofwhich is herein incorporated by reference.

As used here, the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used here, the term “pharmaceutically-acceptable carrier” means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or steric acid), orsolvent encapsulating material. A pharmaceutically acceptable carrierwill not promote the raising of an immune response to an agent withwhich it is admixed, unless so desired. Each carrier must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not injurious to the patient. Some examples ofmaterials which can serve as pharmaceutically-acceptable carriersinclude: (1) sugars, such as lactose, glucose and sucrose; (2) starches,such as corn starch and potato starch; (3) cellulose, and itsderivatives, such as sodium carboxymethyl cellulose, methylcellulose,ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, suchas magnesium stearate, sodium lauryl sulfate and talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12)esters, such as ethyl oleate and ethyl laurate; (13) agar; (14)buffering agents, such as magnesium hydroxide and aluminum hydroxide;(15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18)Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21)polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents,such as polypeptides and amino acids (23) serum component, such as serumalbumin, HDL and LDL; (22) C₂-C₁₂ alcohols, such as ethanol; and (23)other non-toxic compatible substances employed in pharmaceuticalformulations. Wetting agents, coloring agents, release agents, coatingagents, sweetening agents, flavoring agents, perfuming agents,preservative and antioxidants can also be present in the formulation.The terms such as “excipient”, “carrier”, “pharmaceutically acceptablecarrier” or the like are used interchangeably herein.

The amount of a compound or composition described herein that can becombined with a carrier material to produce a single dosage form willgenerally be that amount of the compound that produces a therapeuticeffect. Generally out of one hundred percent, this amount will rangefrom about 0.001% to 99% of the compound, preferably from about 0.01% toabout 70%, most preferably from 5% to about 30%.

Toxicity and therapeutic efficacy can be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀.Compositions that exhibit large therapeutic indices, are preferred.

As used herein, the term ED denotes effective dose and is used inconnection with animal models. The term EC denotes effectiveconcentration and is used in connection with in vitro models.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage may vary within this range depending upon the dosage formemployed and the route of administration utilized.

The therapeutically effective dose can be estimated initially from cellculture assays. A dose may be formulated in animal models to achieve acirculating plasma concentration range that includes the IC₅₀ (i.e., theconcentration of the therapeutic which achieves a half-maximalinhibition of symptoms) as determined in cell culture. Levels in plasmamay be measured, for example, by high performance liquid chromatography.The effects of any particular dosage can be monitored by a suitablebioassay.

The dosage may be determined by a physician and adjusted, as necessary,to suit observed effects of the treatment. Generally, the compositionsare administered so that a compound or composition described herein isgiven at a dose from 1 μg/kg to 150 mg/kg, 1 μg/kg to 100 mg/kg, 1 μg/kgto 50 mg/kg, 1 μg/kg to 20 mg/kg, 1 μg/kg to 10 mg/kg, 1 μg/kg to 1mg/kg, 100 μg/kg to 100 mg/kg, 100 μg/kg to 50 mg/kg, 100 μg/kg to 20mg/kg, 100 μg/kg to 10 mg/kg, 100 μg/kg to 1 mg/kg, 1 mg/kg to 100mg/kg, 1 mg/kg to 50 mg/kg, 1 mg/kg to 20 mg/kg, 1 mg/kg to 10 mg/kg, 10mg/kg to 100 mg/kg, 10 mg/kg to 50 mg/kg, or 10 mg/kg to 20 mg/kg. It isto be understood that ranges given here include all intermediate ranges,for example, the range 1 mg/kg to 10 mg/kg includes 1 mg/kg to 2 mg/kg,1 mg/kg to 3 mg/kg, 1 mg/kg to 4 mg/kg, 1 mg/kg to 5 mg/kg, 1 mg/kg to 6mg/kg, 1 mg/kg to 7 mg/kg, 1 mg/kg to 8 mg/kg, 1 mg/kg to 9 mg/kg, 2mg/kg to 10 mg/kg, 3 mg/kg to 10 mg/kg, 4 mg/kg to 10 mg/kg, 5 mg/kg to10 mg/kg, 6 mg/kg to 10 mg/kg, 7 mg/kg to 10 mg/kg, 8 mg/kg to 10 mg/kg,9 mg/kg to 10 mg/kg, and the like. It is to be further understood thatthe ranges intermediate to the given above are also within the scope ofthis invention, for example, in the range 1 mg/kg to 10 mg/kg, doseranges such as 2 mg/kg to 8 mg/kg, 3 mg/kg to 7 mg/kg, 4 mg/kg to 6mg/kg, and the like.

With respect to duration and frequency of treatment, it is typical forskilled clinicians to monitor subjects in order to determine when thetreatment is providing therapeutic benefit, and to determine whether toincrease or decrease dosage, increase or decrease administrationfrequency, discontinue treatment, resume treatment or make otheralteration to treatment regimen. The dosing schedule can vary from oncea week to daily depending on a number of clinical factors, such as thesubject's sensitivity to the conjugates described herein. The desireddose can be administered everyday or every third, fourth, fifth, orsixth day. The desired dose can be administered at one time or dividedinto subdoses, e.g., 2-4 subdoses and administered over a period oftime, e.g., at appropriate intervals through the day or otherappropriate schedule. Such sub-doses can be administered as unit dosageforms. In some embodiments of the aspects described herein,administration is chronic, e.g., one or more doses daily over a periodof weeks or months. Examples of dosing schedules are administrationdaily, twice daily, three times daily or four or more times daily over aperiod of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3months, 4 months, 5 months, or 6 months or more.

The compounds or the compositions described herein can be administratedto a subject in combination with one or more pharmaceutically activeagents. Exemplary pharmaceutically active compound include, but are notlimited to, those found in Harrison's Principles of Internal Medicine,13^(th) Edition, Eds. T. R. Harrison et al. McGraw-Hill N.Y., NY;Physicians Desk Reference, 50^(th) Edition, 1997, Oradell New Jersey,Medical Economics Co.; Pharmacological Basis of Therapeutics, 8^(th)Edition, Goodman and Gilman, 1990; United States Pharmacopeia, TheNational Formulary, USP XII NF XVII, 1990; current edition of Goodmanand Oilman's The Pharmacological Basis of Therapeutics; and currentedition of The Merck Index, the complete content of all of which areherein incorporated in its entirety.

The compound or the composition and the pharmaceutically active agentcan be administrated to the subject in the same pharmaceuticalcomposition or in different pharmaceutical compositions (at the sametime or at different times). When administrated at different times, theycan be administered within 5 minutes, 10 minutes, 20 minutes, 60minutes, 2 hours, 3 hours, 4, hours, 8 hours, 12 hours, 24 hours ofadministration of the other. When administered in differentpharmaceutical compositions, routes of administration can be different.

Anti-Inflammatory Agents

As used herein, the term “anti-inflammatory compound” or“anti-inflammatory agent” is used to describe any compound (includingits analogs, derivatives, prodrugs and pharmaceutically salts) which maybe used to treat inflammation or inflammation related disease ordisorder. Anti-inflammatory agents include, but are not limited to, theknown steroidal anti-inflammatory and non-steroidal antiinflammatorydrugs (NSAIDs). Exemplary steroidal anti-inflammatory agents include butare not limited to 21-acetoxypregnenolone, alclometasone, algestone,amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone,clobetasol, clobetansone, clocortolone, cloprednol, corticosterone,cortisone, cortivazol, deflazacort, desonide, desoximetasone,dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone,fluazacort, flucloronide, flumethasone flunisolide, fluocinoloneacetonide, fluocinonide, fluocortin butyl, fluocortolone,fluorometholone, fluperolone acetate, fluprednidene acetate,fluprednisolone, flurandrenolide, fluticasone propionate, formocortal,halcinonide, halobetasol propionate, halometasone, halopredone acetate,hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone,medrysone, meprednisone, methylprednisolone, mometasone furcate,paramethosone, prednicarbate, prednisolone, prednisolone25-diethylamino-acetate, prednisolone sodium phosphate, prednisone,prednival, prednylidene, rimexolone, tixocortol, triamcinolone,triamcinolone acetonide, triamcinolone benetonide, triamcinolonehexacetonide, derivatives thereof and mixtures thereof. Exemplarynonsteroidal anti-inflammatory agents include but are not limited to COXinhibitors (COX-1 or COX nonspecific inhibitors) and selective COX-2inhibitors. Exemplary COX inhibitors include but are not limited tosalicylic acid derivatives such as aspirin, sodium salicylate, cholinemagnesium trisalicylate, salicylate, diflunisal, sulfasalazine andolsalazine; para-aminophenol derivatives such as acetaminophen; indoleand indene acetic acids such as indomethacin and sulindac; heteroarylacetic acids such as tolmetin, dicofenac and ketorolac; arylpropionicacids such as ibuprofen, naproxen, flurbiprofen, ketoprofen, fenoprofenand oxaprozin; anthranilic acids (fenamates) such as mefenamic acid andmeloxicam; enolic acids such as the oxicams (piroxicam, meloxicam);alkanones such as nabumetone; derivatives thereof and mixtures thereof.Exemplary COX-2 inhibitors include but are not limited todiarylsubstituted furanones such as refecoxib; diaryl-substitutedpyrazoles such as celecoxib; indole acetic acids such as etodolac andsulfonanilides such as nimesulide; derivatives thereof and mixturesthereof.

Anti-Cancer Agents

As used herein, the term “anti-cancer compound” or “anti-cancer agent”is used to describe any compound (including its analogs, derivatives,prodrugs and pharmaceutically salts) which may be used to treat cancer.Anti-cancer compounds for use in the present invention include, but arenot limited to, inhibitors of topoisomerase I and II, alkylating agents,microtubule inhibitors (e.g., taxol), and angiogenesis inhibitors.Exemplary anti-cancer compounds include, but are not limited to,paclitaxel (taxol); docetaxel; germicitibine; Aldesleukin; Alemtuzumab;alitretinoin; allopurinol; altretamine; amifostine; anastrozole; arsenictrioxide; Asparaginase; BCG Live; bexarotene capsules; bexarotene gel;bleomycin; busulfan intravenous; busulfanoral; calusterone;capecitabine; carboplatin; carmustine; carmustine with PolifeprosanImplant; celecoxib; chlorambucil; cisplatin; cladribine;cyclophosphamide; cytarabine; cytarabine liposomal; dacarbazine;dactinomycin; actinomycin D; Darbepoetin alfa; daunorubicin liposomal;daunorubicin, daunomycin; Denileukin diftitox, dexrazoxane; docetaxel;doxorubicin; doxorubicin liposomal; Dromostanolone propionate; Elliott'sB Solution; epirubicin; Epoetin alfa estramustine; etoposide phosphate;etoposide (VP-16); exemestane; Filgrastim; floxuridine (intraarterial);fludarabine; fluorouracil (5-FU); fulvestrant; gemtuzumab ozogamicin;goserelin acetate; hydroxyurea; Ibritumomab Tiuxetan; idarubicin;ifosfamide; imatinib mesylate; Interferon alfa-2a; Interferon alfa-2b;irinotecan; letrozole; leucovorin; levamisole; lomustine (CCNU);mechlorethamine (nitrogenmustard); megestrol acetate; melphalan (L-PAM);mercaptopurine (6-MP); mesna; methotrexate; methoxsalen; mitomycin C;mitotane; mitoxantrone; nandrolone phenpropionate; Nofetumomab; LOddC;Oprelvekin; oxaliplatin; pamidronate; pegademase; Pegaspargase;Pegfilgrastim; pentostatin; pipobroman; plicamycin; mithramycin;porfimer sodium; procarbazine; quinacrine; Rasburicase; Rituximab;Sargramostim; streptozocin; talbuvidine (LDT); talc; tamoxifen;temozolomide; teniposide (VM-26); testolactone; thioguanine (6-TG);thiotepa; topotecan; toremifene; Tositumomab; Trastuzumab; tretinoin(ATRA); Uracil Mustard; valrubicin; valtorcitabine (monoval LDC);vinblastine; vinorelbine; zoledronate; and any mixtures thereof. In someembodiments, the anti-cancer agent is a paclitaxel-carbohydrateconjugate, e.g., a paclitaxel-glucose conjugate, as described in U.S.Pat. No. 6,218,367, content of which is herein incorporated by referencein its entirety. Some exemplary paclitaxel-carbohydrate conjugatesinclude, but are not limited to, 2′-(GABA-succinoyl)paclitaxel,2′-(glucose-GABA-succinoyl)paclitaxel, 2′-(glucose-succinoyl)paclitaxel,2′-(glucose-glutamyl)paclitaxel,2′-(glucosamide-GABA-succinoyl)paclitaxel,2′-(glucosamide-succinoyl)paclitaxel,2′-(glucosamide-glutamyl)paclitaxel, 7-(GABA-succinoyl)paclitaxel,7-(glucose-GABA-succinoyl)paclitaxel, 7-(glucose-succinoyl)paclitaxel,7-(glucose-glutamyl)paclitaxel,7-(glucosamide-GABA-succinoyl)paclitaxel,7-(glucosamide-succinoyl)paclitaxel, and7-(glucosamide-glutamyl)paclitaxel.

DEFINITIONS

Unless otherwise defined herein, scientific and technical terms used inconnection with the present application shall have the meanings that arecommonly understood by those of ordinary skill in the art. Further,unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular.

As used herein the term “comprising” or “comprises” is used in referenceto compositions, methods, and respective component(s) thereof, that areessential to the invention, yet open to the inclusion of unspecifiedelements, whether essential or not.

As used herein the term “consisting essentially of” refers to thoseelements required for a given embodiment. The term permits the presenceof additional elements that do not materially affect the basic and novelor functional characteristic(s) of that embodiment of the invention.

The term “consisting of” refers to compositions, methods, and respectivecomponents thereof as described herein, which are exclusive of anyelement not recited in that description of the embodiment.

Other than in the operating examples, or where otherwise indicated, allnumbers expressing quantities of ingredients or reaction conditions usedherein should be understood as modified in all instances by the term“about.” The term “about” when used in connection with percentages maymean ±1%. Furthermore, the term “about” can mean within ±1% of a value.

The singular terms “a,” “an,” and “the” include plural referents unlesscontext clearly indicates otherwise. Similarly, the word “or” isintended to include “and” unless the context clearly indicatesotherwise. It is further to be understood that all base sizes or aminoacid sizes, and all molecular weight or molecular mass values, given fornucleic acids or polypeptides are approximate, and are provided fordescription.

Although methods and materials similar or equivalent to those describedherein can be used in the practice or testing of this disclosure,suitable methods and materials are described below. The abbreviation,“e.g.” is derived from the Latin exempli gratia, and is used herein toindicate a non-limiting example. Thus, the abbreviation “e.g.” issynonymous with the term “for example.”

The terms “decrease”, “reduced”, “reduction”, “decrease” or “inhibit”are all used herein generally to mean a decrease by a statisticallysignificant amount. However, for avoidance of doubt, “reduced”,“reduction” or “decrease” or “inhibit” means a decrease by at least 10%as compared to a reference level, for example a decrease by at leastabout 20%, or at least about 30%, or at least about 40%, or at leastabout 50%, or at least about 60%, or at least about 70%, or at leastabout 80%, or at least about 90% or up to and including a 100% decrease(e.g. absent level as compared to a reference sample), or any decreasebetween 10-100% as compared to a reference level.

The terms “increased”,“increase” or “enhance” or “activate” are all usedherein to generally mean an increase by a statically significant amount;for the avoidance of any doubt, the terms “increased”, “increase” or“enhance” or “activate” means an increase of at least 10% as compared toa reference level, for example an increase of at least about 20%, or atleast about 30%, or at least about 40%, or at least about 50%, or atleast about 60%, or at least about 70%, or at least about 80%, or atleast about 90% or up to and including a 100% increase or any increasebetween 10-100% as compared to a reference level, or at least about a2-fold, or at least about a 3-fold, or at least about a 4-fold, or atleast about a 5-fold or at least about a 10-fold increase, or anyincrease between 2-fold and 10-fold or greater as compared to areference level.

The term “statistically significant” or “significantly” refers tostatistical significance and generally means a two standard deviation(2SD) above or below a reference level. The term refers to statisticalevidence that there is a difference. It is defined as the probability ofmaking a decision to reject the null hypothesis when the null hypothesisis actually true. The decision is often made using the p-value.

As used herein, the term “synergistic” means a combination of componentswherein the activity of the combination is greater than the additive ofthe individual activities of each component of the combination

As used herein, the term “anti-cancer activity” or “anti-cancerproperties” refers to the inhibition (in part or in whole) or preventionof unregulated cell growth and/or the inhibition (in part or in whole)or prevention of a cancer as defined herein. Anticancer activityincludes, e.g., the ability to reduce, prevent, or repair geneticdamage, modulate undesired cell proliferation, modulate misregulatedcell death, or modulate mechanisms of metastasis (e.g., ability tomigrate).

As used herein, the term “anti-inflammatory activity” refers toprevention or reduction of one or more indicia of inflammation.

As used herein, the term “analgesic” refers to a compound capable ofproducing analgesia, i.e., reducing or inhibiting pain by alteringperception of nociceptive stimuli without producing anesthesia or lossof consciousness.

By “treatment”, “prevention” or “amelioration” of a disease or disorderis meant delaying or preventing the onset of such a disease or disorder,reversing, alleviating, ameliorating, inhibiting, slowing down orstopping the progression, aggravation or deterioration the progressionor severity of a condition associated with such a disease or disorder.In one embodiment, one or more symptoms of a disease or disorder arealleviated by at least 5%, at least 10%, at least 20%, at least 30%, atleast 40%, or at least 50%.

As used herein, the term “oligosaccharide” refers without limitation toseveral (e.g., two to ten) covalently linked monosaccharide units.Oligosaccharides include, but are not limited to, disaccharides (i.e.,two monosaccharide units) such as sucrose, lactose, maltose, isomaltose,cellobiose and the like.

As used herein, the term “polysaccharide” refers without limitation tomany (e.g., eleven or more) covalently linked monosaccharide units.Polysaccharides can have molecular masses ranging well into millions ofdaltons. The polysaccharide can be homopolysaccharides orheteropolysaccharides. Whereas the homopolysaccharides contain only onekind of unit, the heteropolysaccharides consist of monomer units ofdifferent kinds. Exemplary polysaccharides include, but are not limitedto, cellulose, chitin, starch, glycogen, glycosaminoglycans (e.g.,hyaluronic acid, chondroitin-4-sulfate, chondroitin-6-sulfate, dermatansulfate, keratin sulfate, heparin and the like) and the like. The di-,tri-, oligo- and polysaccharides can comprise 1->4, 1->6 or a mixture of1->4 and 1->6 linkages.

As used herein, the term “fish oil” indicates oil and fat extracted fromanimals living in water (namely, fish oil in a broad sense) and fish oilextracted from fish (fish oil in a narrow sense), sea animals oil andcod-liver oil are included. All type of said fish oil (fish oil in broadsense) contains EPA, DHA and other types of highly unsaturated fattyacid and/or their esters. And the term “fish oil extract” indicatesfatty acid having 2 or more unsaturated bonds obtained by refining andseparating of said fish oil in broad sense, and as the concrete example,EPA or DHA can be mentioned, however the invention is, not limited tothem. In the present invention, the following can be used as the fishoil: crude fish oil; lower refined fish oil; highly refined fish oil;and mixtures thereof.

In general, fish oil extracted from sardine, mackerel, codfish and tuna(fish oil in a narrow sense), lower and highly refined oil of it, highlyunsaturated fatty acid obtained from said fish oil and its ester arepreferably used. Further, in the present invention, the fish oil or fishoil extract which comprises at least 10% by weight of DHA and/or EPA isdesirable.

The term “alkyl” refers to saturated non-aromatic hydrocarbon chainsthat may be a straight chain or branched chain, containing the indicatednumber of carbon atoms (these include without limitation methyl, ethyl,propyl, iso-propyl, butyl, 2-methyl-ethyl, t-butyl, allyl, orpropargyl), which may be optionally inserted with N, O, or S. Forexample, C₁-C₆ indicates that the group may have from 1 to 6 carbonatoms in it.

The term “alkenyl” refers to an alkyl that comprises at least one doublebond. Exemplary alkenyl groups include, but are not limited to, forexample, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl and the like.

The term “alkynyl” refers to an alkyl that comprises at least one triplebond.

The term “cyclyl” or “cycloalkyl” refers to saturated and partiallyunsaturated cyclic hydrocarbon groups having 3 to 12 carbons, forexample, 3 to 8 carbons, and, for example, 3 to 6 carbons, wherein thecycloalkyl group additionally may be optionally substituted. Exemplarycycloalkyl groups include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,cycloheptyl, cyclooctyl, and the like.

The term “heterocyclyl” refers to a nonaromatic 5-8 membered monocyclic,8-12 membered bicyclic, or 11-14 membered tricyclic ring system having1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms selected from O, N, or S(e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S ifmonocyclic, bicyclic, or tricyclic, respectively), wherein 0, 1, 2 or 3atoms of each ring may be substituted by a substituent. Exemplaryheterocyclyl groups include, but are not limited to piperazinyl,pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, and the like.

The term “aryl” refers to monocyclic, bicyclic, or tricyclic aromaticring system wherein 0, 1, 2, 3, or 4 atoms of each ring may besubstituted by a substituent. Exemplary aryl groups include, but are notlimited to, phenyl, naphthyl, anthracenyl, azulenyl, fluorenyl, indanyl,indenyl, naphthyl, phenyl, tetrahydronaphthyl, and the like.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic,8-12 membered bicyclic, or 11-14 membered tricyclic ring system having1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms selected from O, N, or S(e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S ifmonocyclic, bicyclic, or tricyclic, respectively), wherein 0, 1, 2, 3,or 4 atoms of each ring may be substituted by a substituent. Exemplaryheteroaryl groups include, but are not limited to, pyridyl, furyl orfuranyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or thienyl,pyridazinyl, pyrazinyl, quinolinyl, indolyl, thiazolyl, naphthyridinyl,and the like.

The term “acyl” refers to an alkylcarbonyl or arylcarbonyl substituent,wherein the alkyl or aryl portion may be optionally substituted.

The term “optionally substituted” means that the specified group ormoiety, such as an alkyl group, alkenyl group, and the like, isunsubstituted or is substituted with one or more (typically 1-4substituents) independently selected from the group of substituentslisted below in the definition for “substituents” or otherwisespecified.

The term “substituents” refers to a group “substituted” on an alkyl,alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl group atany atom of that group. Suitable substituents include, withoutlimitation, halogen, hydroxy, oxo, nitro, haloalkyl, alkyl, alkenyl,alkynyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino,alkylcarbanoyl, arylcarbanoyl, aminoalkyl, alkoxycarbonyl, carboxy,hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido,arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano orureido. In some cases, two substituents, together with the carbons towhich they are attached to can form a ring.

As used here in the term “isomer” refers to compounds having the samemolecular formula but differing in structure. Isomers which differ onlyin configuration and/or conformation are referred to as “stereoisomers.”The term “isomer” is also used to refer to an enantiomer.

The term “analog” as used herein refers to a compound that results fromsubstitution, replacement or deletion of various organic groups orhydrogen atoms from a parent compound. As such, some monoterpenoids canbe considered to be analogs of monoterpenes, or in some cases, analogsof other monoterpenoids, including derivatives of monoterpenes. Ananalog is structurally similar to the parent compound, but can differ byeven a single element of the same valence and group of the periodictable as the element it replaces.

The term “derivative” as used herein refers to a chemical substancerelated structurally to another, i.e., an “original” substance, whichcan be referred to as a “parent” compound. A “derivative” can be madefrom the structurally-related parent compound in one or more steps. Thephrase “closely related derivative” means a derivative whose molecularweight does not exceed the weight of the parent compound by more than50%. The general physical and chemical properties of a closely relatedderivative are also similar to the parent compound.

As used herein, a “prodrug” refers to compounds that can be convertedvia some chemical or physiological process (e.g., enzymatic processesand metabolic hydrolysis) to a therapeutic agent. Thus, the term“prodrug” also refers to a precursor of a biologically active compoundthat is pharmaceutically acceptable. A prodrug may be inactive whenadministered to a subject, i.e. an ester, but is converted in vivo to anactive compound, for example, by hydrolysis to the free carboxylic acidor free hydroxyl. The prodrug compound often offers advantages ofsolubility, tissue compatibility or delayed release in an organism. Theterm “prodrug” is also meant to include any covalently bonded carriers,which release the active compound in vivo when such prodrug isadministered to a subject. Prodrugs of an active compound may beprepared by modifying functional groups present in the active compoundin such a way that the modifications are cleaved, either in routinemanipulation or in vivo, to the parent active compound. Prodrugs includecompounds wherein a hydroxy, amino or mercapto group is bonded to anygroup that, when the prodrug of the active compound is administered to asubject, cleaves to form a free hydroxy, free amino or free mercaptogroup, respectively. Examples of prodrugs include, but are not limitedto, acetate, formate and benzoate derivatives of an alcohol oracetamide, formamide and benzamide derivatives of an amine functionalgroup in the active compound and the like. See Harper, “DrugLatentiation” in Jucker, ed. Progress in Drug Research 4:221-294 (1962);Morozowich et al, “Application of Physical Organic Principles to ProdrugDesign” in E. B. Roche ed. Design of Biopharmaceutical Propertiesthrough Prodrugs and Analogs, APHA Acad. Pharm. Sci. 40 (1977);Bioreversible Carriers in Drug in Drug Design, Theory and Application,E. B. Roche, ed., APHA Acad. Pharm. Sci. (1987); Design of Prodrugs, H.Bundgaard, Elsevier (1985); Wang et al. “Prodrug approaches to theimproved delivery of peptide drug” in Curr. Pharm. Design. 5(4):265-287(1999); Pauletti et al. (1997) Improvement in peptide bioavailability:Peptidomimetics and Prodrug Strategies, Adv. Drug. Delivery Rev.27:235-256; Mizen et al. (1998) “The Use of Esters as Prodrugs for OralDelivery of (3-Lactam antibiotics,” Pharm. Biotech. 11:345-365;Gaignault et al. (1996) “Designing Prodrugs and Bioprecursors I. CarrierProdrugs,” Pract. Med. Chem. 671-696; Asgharnejad, “Improving Oral DrugTransport”, in Transport Processes in Pharmaceutical Systems, G. L.Amidon, P. I. Lee and E. M. Topp, Eds., Marcell Dekker, p. 185-218(2000); Balant et al., “Prodrugs for the improvement of drug absorptionvia different routes of administration”, Eur. J. Drug Metab.Pharmacokinet., 15(2): 143-53 (1990); Balimane and Sinko, “Involvementof multiple transporters in the oral absorption of nucleosideanalogues”, Adv. Drug Delivery Rev., 39(1-3): 183-209 (1999); Browne,“Fosphenytoin (Cerebyx)”, Clin. Neuropharmacol. 20(1): 1-12 (1997);Bundgaard, “Bioreversible derivatization of drugs—principle andapplicability to improve the therapeutic effects of drugs”, Arch. Pharm.Chemi 86(1): 1-39 (1979); Bundgaard H. “Improved drug delivery by theprodrug approach”, Controlled Drug Delivery 17: 179-96 (1987); BundgaardH. “Prodrugs as a means to improve the delivery of peptide drugs”, Arfv.Drug Delivery Rev. 8(1): 1-38 (1992); Fleisher et al. “Improved oraldrug delivery: solubility limitations overcome by the use of prodrugs”,Arfv. Drug Delivery Rev. 19(2): 115-130 (1996); Fleisher et al. “Designof prodrugs for improved gastrointestinal absorption by intestinalenzyme targeting”, Methods Enzymol. 112 (Drug Enzyme Targeting, Pt. A):360-81, (1985); Farquhar D, et al., “Biologically ReversiblePhosphate-Protective Groups”, Pharm. Sci., 72(3): 324-325 (1983);Freeman S, et al., “Bioreversible Protection for the Phospho Group:Chemical Stability and Bioactivation of Di(4-acetoxy-benzyl)Methylphosphonate with Carboxyesterase,” Chem. Soc., Chem. Commun.,875-877 (1991); Friis and Bundgaard, “Prodrugs of phosphates andphosphonates: Novel lipophilic alphaacyloxyalkyl ester derivatives ofphosphate- or phosphonate containing drugs masking the negative chargesof these groups”, Eur. J. Pharm. Sci. 4: 49-59 (1996); Gangwar et al.,“Pro-drug, molecular structure and percutaneous delivery”, Des.Biopharm. Prop. Prodrugs Analogs, [Symp.] Meeting Date 1976, 409-21.(1977); Nathwani and Wood, “Penicillins: a current review of theirclinical pharmacology and therapeutic use”, Drugs 45(6): 866-94 (1993);Sinhababu and Thakker, “Prodrugs of anticancer agents”, Adv. DrugDelivery Rev. 19(2): 241-273 (1996); Stella et al., “Prodrugs. Do theyhave advantages in clinical practice?”, Drugs 29(5): 455-73 (1985); Tanet al. “Development and optimization of anti-HIV nucleoside analogs andprodrugs: A review of their cellular pharmacology, structure-activityrelationships and pharmacokinetics”, Adv. Drug Delivery Rev. 39(1-3):117-151 (1999); Taylor, “Improved passive oral drug delivery viaprodrugs”, Adv. Drug Delivery Rev., 19(2): 131-148 (1996); Valentino andBorchardt, “Prodrug strategies to enhance the intestinal absorption ofpeptides”, Drug Discovery Today 2(4): 148-155 (1997); Wiebe and Knaus,“Concepts for the design of anti-HIV nucleoside prodrugs for treatingcephalic HIV infection”, Adv. Drug Delivery Rev.: 39(1-3):63-80 (1999);Waller et al., “Prodrugs”, Br. J. Clin. Pharmac. 28: 497-507 (1989),content of all of which is herein incorporated by reference in itsentirety.

As used herein, the term “pharmaceutically-acceptable salts” refers tothe conventional nontoxic salts or quaternary ammonium salts oftherapeutic agents, e.g., from non-toxic organic or inorganic acids.These salts can be prepared in situ in the administration vehicle or thedosage form manufacturing process, or by separately reacting atherapeutic agent in its free base or acid form with a suitable organicor inorganic acid or base, and isolating the salt thus formed duringsubsequent purification. Conventional nontoxic salts include thosederived from inorganic acids such as sulfuric, sulfamic, phosphoric,nitric, and the like; and the salts prepared from organic acids such asacetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric,citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic,glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic,and the like. See, for example, Berge et al., “Pharmaceutical Salts”, J.Pharm. Sci. 66:1-19 (1977), content of which is herein incorporated byreference in its entirety.

In some embodiments of the aspects described herein, representativesalts include the hydrobromide, hydrochloride, sulfate, bisulfate,phosphate, nitrate, acetate, succinate, valerate, oleate, palmitate,stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate,maleate, fumarate, succinate, tartrate, naphthylate, mesylate,glucoheptonate, lactobionate, and laurylsulphonate salts and the like.

The present invention can be defined in any of the following numberedparagraphs:

-   1. A turmeric oil extract obtained by high vacuum distillation of    turmeric oil and collecting a distillate at 70-100° C. or at    100-130° C.-   2. The turmeric oil extract of paragraph 1, wherein the extract is    produced by a process comprising the steps of: (i) extracting a    turmeric powder with hexane; (ii) distilling the extract of (i) to    obtain a distillate at 115-135° C. under high vacuum; (iii)    distilling the distillate of (ii) to obtain a distillate at    95-112° C. under high vacuum; (iv) distilling the distillate    of (iii) to obtain a distillate at 100-110° C. under high vacuum;    and (v) distilling the distillate of (iv) to obtain the extract as a    distillate at 120-123° C. under high vacuum.-   3. The turmeric oil extract of paragraph 1, wherein the extract is    produced by a process comprising the steps of: (i) extracting a    turmeric powder with hexane; (ii) distilling the extract of (i) to    obtain a distillate at 115-135° C. under high vacuum; (iii)    distilling the distillate of (ii) to obtain a distillate at    95-112° C. under high vacuum; (iv) distilling the distillate    of (iii) to obtain a distillate at 100-110° C. under high    vacuum; (v) distilling the distillate of (iv) to obtain a distillate    at 100-120° C. and at 124° C. under high vacuum; (vi) combining the    distillates obtained in (v) and obtaining the extract by eluting the    combined distillates from a column using one volume of hexane, one    volume 5% of ethyl acetate/hexane, and one volume 1% ethyl    acetate/Hexane.-   4. The turmeric oil extract of any of paragraphs 1-3, wherein the    high vacuum is less than about 250 torr.-   5. The turmeric oil extract of any of paragraphs 1-4, wherein at    least one compound in the extract comprises at least 50% of the    extract.-   6. The turmeric oil extract of any of paragraphs 1-5, wherein the    extract comprises from about 70 to about 75% carbon and from about 5    to about 10% hydrogen.-   7. The turmeric oil extract of any of paragraphs 1-6, wherein the    extract has NMR spectra shown in FIG. 2A, FIG. 2B, FIG. 3A, FIG. 3B,    or FIG. 3C.-   8. The turmeric oil extract of any of paragraphs 1-7, wherein the    extract is anti-inflammatory, anti-cancer and/or analgesic.-   9. The turmeric oil extract of any of paragraphs 1-8, wherein the    extract has a synergistic anti-inflammatory activity and/or    analgesic activity with an anti-inflammatory agent, or the extract    has a synergistic anti-cancer activity with an anti-cancer agent.-   10. The turmeric oil extract of any of paragraphs 1-9, wherein the    extract enhances anti-inflammatory activity of an anti-inflammatory    agent or the extract enhances anti-cancer activity of an anti-cancer    agent.-   11. A curcumin derivative having the structure of formula (I):    -   wherein:

-   -   R¹ and R² are independently H, optionally substituted alkyl,        optionally substituted alkenyl, optionally substituted alkynyl,        optionally substituted cyclyl, optionally substituted        heterocyclyl, optionally substituted aryl, optionally        substituted heteroaryl, a peptide, —C(O)R³, —C(O)OR³, or        —C(O)NR³R³, provided that at least one of R¹ and R² is not H;    -   R³ is independently for each occurrence H, optionally        substituted alkyl, optionally substituted alkenyl, optionally        substituted alkynyl, optionally substituted cyclyl, optionally        substituted heterocyclyl, optionally substituted aryl,        optionally substituted heteroaryl; and analogs, derivatives,        isomers, prodrugs, and pharmaceutically acceptable salts        thereof.

-   12. The curcumin derivative of paragraph 11, wherein R¹ and R² are    the same.

-   13. The curcumin derivative of paragraph 11, wherein one of R¹ and    R² is H.

-   14. The curcumin derivative of any of paragraphs 11-13, wherein at    least one of R¹ and R² is selected from the group consisting of    acetyl, myristoleoyl, palmitoleoyl, sapienoyl, oleoyl, linoleoyl,    α-linoleoyl, α-linolenoyl, γ-linolenoyl, arcchidionoyl,    eicosapentaenoyl, erucoyl, docosahexaenoyl, lauroyl, myrsitoyl,    palmitoyl, stearoyl, arachidoyl, behenoyl, lignoceroyl, certoyl and    any combinations thereof.

-   15. The curcumin derivative of any of paragraphs 11-14, wherein at    least one of R¹ and R² is —C(O)R³ and R³ is an optionally    substituted aryl.

-   16. The curcumin derivative of paragraph 15, wherein the aryl is    substituted at the 2-, 3-, 4-, or 5-position or any combinations of    these positions.

-   17. The curcumin derivative of any of paragraphs 15-16, wherein the    optionally substituted aryl is an optionally substituted phenyl

-   18. The curcumin derivative of any of paragraphs 11-17, wherein at    least one of R¹ and R² is

-    wherein R⁴ is H, optionally substituted alkyl, optionally    substituted alkenyl, optionally substituted alkynyl, or optionally    substituted acyl.-   19. The curcumin derivative of paragraph 18, wherein R⁴ is H or    C(O)CH₃.-   20. The curcumin derivative of any of paragraphs 11-19, wherein at    least one of R¹ and R² is -linker-R⁵, wherein R⁵ is H, a    carbohydrate, a peptide, and analogs and derivatives thereof.-   21. The curcumin derivative of paragraph 20, wherein the linker is    —C(O)(CH₂)_(m)C(O)O—, wherein m is an integer from 1 to 10.-   22. The curcumin derivative of paragraph 21, wherein m is 2 or 3-   23. The curcumin derivative of any of paragraphs 20-22, wherein R⁵    is H, a carbohydrate or a peptide.-   24. The curcumin derivative of paragraph 11, wherein R¹ and R² both    are not —C(O)CH₃.-   25. The curcumin derivative of paragraph 11, wherein the curcumin    derivative is di(acetylsalicyloyl)-curcumin    monoacetylsalicyloyl-curcumin, diacetyl-curcumin,    monoacetyl-curcumin, diaglutaroyl-curcumin, monoglutaroyl-curcumin,    di-gluocose-glutaroyl-curcumin, mono-gluocose-glutaroyl-curcumin,    monolinoleol-curcumnin, di-linoleoyl-curcumin and peptide-curcumin    conjugates.-   26. The curcumin derivative of any of paragraph 11-25, wherein the    curcumin derivative is anti-inflammatory, anti-cancer, and/or    analgesic.-   27. The curcumin derivative of any of paragraph 11-26, wherein the    curcumin derivative has synergistic anti-inflammatory activity    and/or analgesic activity with an anti-inflammatory agent or the    curcumin derivative has synergistic anti-cancer activity with an    anti-cancer agent.-   28. The curcumin derivative of any of paragraph 11-27, wherein the    curcumin derivative enhances anti-inflammatory activity of an    anti-inflammatory agent or the curcumin derivative enhances    anti-cancer activity of an anti-cancer agent.-   29. A composition comprising turmeric oil or a turmeric oil extract    and a compound selected from the group consisting of an anti-cancer    compound, an anti-inflammatory agent, a curcumin conjugate of any of    paragraphs 11-28, fish oil, fish oil extract, a salicylic acid    conjugate, a curcumin ether derivative, and any combinations    thereof, wherein the salicylic acid is of formula (II):

-   -   wherein R⁸ is a carbohydrate; R⁹ is H, optionally substituted        alkyl, optionally substituted alkenyl, optionally substituted        alkynyl, or optionally substituted acyl; and analogs,        derivatives, isomers, prodrugs, and pharmaceutically acceptable        salts thereof,    -   and wherein the curcumin ether derivative is of formula (III):

-   -   wherein R₂ and R₄ are both CH₃; and R₁ and R₃ are both alkyl or        one of R₁ and R₃ is H and the other is alkyl.

-   30. The composition of paragraph 29, wherein the composition    comprises at least two of: (a) an anti-cancer compound; (b) a    curcumin derivative; (c) fish oil or fish oil extract; (e) a    salicylic acid conjugate; and (f) a curcumin ether derivative.

-   31. The composition of paragraph 30, wherein the composition    comprises salicylic conjugate and a curcumin derivative.

-   32. The composition of any of paragraphs 29-31, wherein the turmeric    oil extract is an extract of any of paragraphs 1-10.

-   33. The composition of any of paragraphs 29-32, wherein R⁸ is    selected from the group consisting of glucose, glyceraldehydes,    erythrose, threose, ribulose, xylulose, ribose, arabinose,    deoxyribose, xylose, lyxose, psicose, fructose, sorbose, tagatose,    allose, altrose, mannose, gulose, idose, galactose, talose, fucose,    fuculose, rhamnose, sedoheptulose, octose, nonose (Neuraminic acid),    and the like.

-   34. The composition of any of paragraphs 29-33, wherein R⁹ is H or    C(O)CH₃.

-   35. The composition of any of paragraphs 29-34, wherein the    salicylic acid conjugate is of formula (IIb):

-   36. The composition of any of paragraphs 29-35, wherein the    anti-cancer agent is selected from the group consisting of    paclitaxel (taxol); docetaxel; germicitibine; Aldesleukin;    Alemtuzumab; alitretinoin; allopurinol; altretamine; amifostine;    anastrozole; arsenic trioxide; Asparaginase; BCG Live; bexarotene    capsules; bexarotene gel; bleomycin; busulfan intravenous;    busulfanoral; calusterone; capecitabine; carboplatin; carmustine;    carmustine with Polifeprosan Implant; celecoxib; chlorambucil;    cisplatin; cladribine; cyclophosphamide; cytarabine; cytarabine    liposomal; dacarbazine; dactinomycin; actinomycin D; Darbepoetin    alfa; daunorubicin liposomal; daunorubicin, daunomycin; Denileukin    diftitox, dexrazoxane; docetaxel; doxorubicin; doxorubicin    liposomal; Dromostanolone propionate; Elliott's B Solution;    epirubicin; Epoetin alfa estramustine; etoposide phosphate;    etoposide (VP-16); exemestane; Filgrastim; floxuridine    (intraarterial); fludarabine; fluorouracil (5-FU); fulvestrant;    gemtuzumab ozogamicin; goserelin acetate; hydroxyurea; Ibritumomab    Tiuxetan; idarubicin; ifosfamide; imatinib mesylate; Interferon    alfa-2a; Interferon alfa-2b; irinotecan; letrozole; leucovorin;    levamisole; lomustine (CCNU); mechlorethamine (nitrogenmustard);    megestrol acetate; melphalan (L-PAM); mercaptopurine (6-MP); mesna;    methotrexate; methoxsalen; mitomycin C; mitotane; mitoxantrone;    nandrolone phenpropionate; Nofetumomab; LOddC; Oprelvekin;    oxaliplatin;    -   pamidronate; pegademase; Pegaspargase; Pegfilgrastim;        pentostatin; pipobroman; plicamycin; mithramycin; porfimer        sodium; procarbazine; quinacrine; Rasburicase; Rituximab;        Sargramostim; streptozocin; talbuvidine (LDT); talc; tamoxifen;        temozolomide; teniposide (VM-26); testolactone; thioguanine        (6-TG); thiotepa; topotecan; toremifene; Tositumomab;        Trastuzumab; tretinoin (ATRA); Uracil Mustard; valrubicin;        valtorcitabine (monoval LDC); vinblastine; vinorelbine;        zoledronate; 2′-(GABA-succinoyl)paclitaxel,        2′-(glucose-GABA-succinoyl)paclitaxel,        2′-(glucose-succinoyl)paclitaxel,        2′-(glucose-glutamyl)paclitaxel,        2′-(glucosamide-GABA-succinoyl)paclitaxel,        2′-(glucosamide-succinoyl)paclitaxel,        2′-(glucosamide-glutamyl)paclitaxel,        7-(GABA-succinoyl)paclitaxel,        7-(glucose-GABA-succinoyl)paclitaxel,        7-(glucose-succinoyl)paclitaxel, 7-(glucose-glutamyl)paclitaxel,        7-(glucosamide-GABA-succinoyl)paclitaxel,        7-(glucosamide-succinoyl)paclitaxel, and        7-(glucosamide-glutamyl)paclitaxel; and any mixtures thereof.-   37. The composition of any of paragraphs 29-36, wherein the    anti-inflammatory agent is selected from the group consisting of    21-acetoxypregnenolone, alclometasone, algestone, amcinonide,    beclomethasone, betamethasone, budesonide, chloroprednisone,    clobetasol, clobetansone, clocortolone, cloprednol, corticosterone,    cortisone, cortivazol, deflazacort, desonide, desoximetasone,    dexamethasone, diflorasone, diflucortolone, difluprednate,    enoxolone, fluazacort, flucloronide, flumethasone flunisolide,    fluocinolone acetonide, fluocinonide, fluocortin butyl,    fluocortolone, fluorometholone, fluperolone acetate, fluprednidene    acetate, fluprednisolone, flurandrenolide, fluticasone propionate,    formocortal, halcinonide, halobetasol propionate, halometasone,    halopredone acetate, hydrocortamate, hydrocortisone, loteprednol    etabonate, mazipredone, medrysone, meprednisone, methylprednisolone,    mometasone furcate, paramethosone, prednicarbate, prednisolone,    prednisolone 25-diethylamino-acetate, prednisolone sodium phosphate,    prednisone, prednival, prednylidene, rimexolone, tixocortol,    triamcinolone, triamcinolone acetonide, triamcinolone benetonide,    triamcinolone hexacetonide, aspirin, sodium salicylate, choline    magnesium trisalicylate, salicylate, diflunisal, sulfasalazine,    olsalazine, acetaminophen, indomethacin, sulindac, tolmetin,    dicofenac, ketorolac, ibuprofen, naproxen, flurbiprofen, ketoprofen,    fenoprofen, oxaprozin, mefenamic acid, meloxicam, oxicams    (piroxicam, meloxicam); alkanones such as nabumetone, refecoxib,    celecoxib, etodolac, sulfonanilides, and derivatives thereof and    mixtures thereof.-   38. The composition of any of paragraphs 29-37, wherein the ratio of    the turmeric oil or the turmeric oil extract and the compound is    from about 99:1 to about 1:99.-   39. The composition of any of paragraphs 29-38, wherein the    composition comprises at least 50% turmeric oil and/or turmeric oil    extract.-   40. The composition of any of paragraphs 29-39, wherein the    composition comprises turmeric oil extract and aspirin; turmeric oil    extract and a curcumin derivative, turmeric oil extract and a    salicylic acid conjugate; turmeric oil extract and    di(acetylsalicyloyl)-curcumin; turmeric oil extract and    monoacetylsalicyloyl-curcumin; turmeric oil extract and    diacetyl-curcumin; turmeric oil extract and monoacetyl-curcumin;    turmeric oil extract and diaglutaroyl-curcumin; turmeric oil extract    and monoglutaroyl-curcumin; turmeric oil extract and    di-gluocose-glutaroyl-curcumin; turmeric oil extract and    mono-gluocose-glutaroyl-curcumin; turmeric oil extract and    monolinoleol-curcumnin; turmeric oil extract and    di-linoleoyl-curcumin; turmeric oil extract and an anticancer agent;    turmeric oil extract and an anti-inflammatory agent; turmeric oil    extract and fish oil; turmeric oil extract and fish oil extract;    turmeric oil extract, an anti-cancer agent and an anti-inflammatory    agent; turmeric oil extract, an anti-cancer agent and a curcumin    derivative; turmeric oil extract, an anti-cancer agent and a    salicylic acid conjugate; turmeric oil extract, an anti-inflammatory    agent and a curcumin derivative; turmeric oil extract, an    anti-inflammatory agent and a salicylic acid conjugate; or turmeric    oil extract, a curcumin derivative and a salicylic acid conjugate.-   41. A composition comprising a curcumin derivative of any of    paragraphs 11-29 and a compound selected from the group consisting    of an anti-cancer compound, an anti-inflammatory agent, fish oil,    fish oil extract, a salicylic acid conjugate, a curcumin ether    derivative, and any combinations thereto, wherein the salicylic acid    conjugate is of formula (II):

-   -   wherein R⁸ is a carbohydrate; R⁹ is H, optionally substituted        alkyl, optionally substituted alkenyl, optionally substituted        alkynyl, or optionally substituted acyl; and analogs,        derivatives, isomers, prodrugs, and pharmaceutically acceptable        salts thereof,    -   and wherein the curcumin ether derivative is of formula (III):

-   -   wherein R₂ and R₄ are both CH₃; and R₁ and R₃ are both alkyl or        one of R₁ and R₃ is H and the other is alkyl.

-   42. The composition of paragraph 41, wherein the composition    comprises at least two of: (a) an anti-cancer compound; (b) fish oil    or fish oil extract; (c) a salicylic acid conjugate; and (e) a    curcumin ether derivative.

-   43. The composition of any of paragraphs 41-42, wherein R⁸ is    selected from the group consisting of glucose, glyceraldehydes,    erythrose, threose, ribulose, xylulose, ribose, arabinose,    deoxyribose, xylose, lyxose, psicose, fructose, sorbose, tagatose,    allose, altrose, mannose, gulose, idose, galactose, talose, fucose,    fuculose, rhamnose, sedoheptulose, octose, nonose (Neuraminic acid),    and the like.

-   44. The composition of any of paragraphs 41-43, wherein R⁹ is H or    C(O)CH₃.

-   45. The composition of any of paragraphs 41-44, wherein the    salicylic acid conjugate is of formula (IIb):

-   46. The composition of any of paragraphs 41-45, wherein the    anti-cancer agent is selected from the group consisting of    paclitaxel (taxol); docetaxal; germicitibine; Aldesleukin;    Alemtuzumab; alitretinoin; allopurinol; altretamine; amifostine;    anastrozole; arsenic trioxide; Asparaginase; BCG Live; bexarotene    capsules; bexarotene gel; bleomycin; busulfan intravenous;    busulfanoral; calusterone; capecitabine; carboplatin; carmustine;    carmustine with Polifeprosan Implant; celecoxib; chlorambucil;    cisplatin; cladribine; cyclophosphamide; cytarabine; cytarabine    liposomal; dacarbazine; dactinomycin; actinomycin D; Darbepoetin    alfa; daunorubicin liposomal; daunorubicin, daunomycin; Denileukin    diftitox, dexrazoxane; docetaxel; doxorubicin; doxorubicin    liposomal; Dromostanolone propionate; Elliott's B Solution;    epirubicin; Epoetin alfa estramustine; etoposide phosphate;    etoposide (VP-16); exemestane; Filgrastim; floxuridine    (intraarterial); fludarabine; fluorouracil (5-FU); fulvestrant;    gemtuzumab ozogamicin; goserelin acetate; hydroxyurea; Ibritumomab    Tiuxetan; idarubicin; ifosfamide; imatinib mesylate; Interferon    alfa-2a; Interferon alfa-2b; irinotecan; letrozole; leucovorin;    levamisole; lomustine (CCNU); mechlorethamine (nitrogenmustard);    megestrol acetate; melphalan (L-PAM); mercaptopurine (6-MP); mesna;    methotrexate; methoxsalen; mitomycin C; mitotane; mitoxantrone;    nandrolone phenpropionate; Nofetumomab; LOddC; Oprelvekin;    oxaliplatin; pamidronate; pegademase; Pegaspargase; Pegfilgrastim;    pentostatin; pipobroman; plicamycin; mithramycin; porfimer sodium;    procarbazine; quinacrine; Rasburicase; Rituximab; Sargramostim;    streptozocin; talbuvidine (LDT); talc; tamoxifen; temozolomide;    teniposide (VM-26); testolactone; thioguanine (6-TG); thiotepa;    topotecan; toremifene; Tositumomab; Trastuzumab; tretinoin (ATRA);    Uracil Mustard; valrubicin; valtorcitabine (monoval LDC);    vinblastine; vinorelbine; zoledronate; and any mixtures thereof.-   47. The composition of any of paragraphs 41-46, wherein the    anti-inflammatory agent is selected from the group consisting of    21-acetoxypregnenolone, alclometasone, algestone, amcinonide,    beclomethasone, betamethasone, budesonide, chloroprednisone,    clobetasol, clobetansone, clocortolone, cloprednol, corticosterone,    cortisone, cortivazol, deflazacort, desonide, desoximetasone,    dexamethasone, diflorasone, diflucortolone, difluprednate,    enoxolone, fluazacort, flucloronide, flumethasone flunisolide,    fluocinolone acetonide, fluocinonide, fluocortin butyl,    fluocortolone, fluorometholone, fluperolone acetate, fluprednidene    acetate, fluprednisolone, flurandrenolide, fluticasone propionate,    formocortal, halcinonide, halobetasol propionate, halometasone,    halopredone acetate, hydrocortamate, hydrocortisone, loteprednol    etabonate, mazipredone, medrysone, meprednisone, methylprednisolone,    mometasone furcate, paramethosone, prednicarbate, prednisolone,    prednisolone 25-diethylamino-acetate, prednisolone sodium phosphate,    prednisone, prednival, prednylidene, rimexolone, tixocortol,    triamcinolone, triamcinolone acetonide, triamcinolone benetonide,    triamcinolone hexacetonide, aspirin, sodium salicylate, choline    magnesium trisalicylate, salicylate, diflunisal, sulfasalazine,    olsalazine, acetaminophen, indomethacin, sulindac, tolmetin,    dicofenac, ketorolac, ibuprofen, naproxen, flurbiprofen, ketoprofen,    fenoprofen, oxaprozin, mefenamic acid, meloxicam, oxicams    (piroxicam, meloxicam); alkanones such as nabumetone, refecoxib,    celecoxib, etodolac, sulfonanilides, and derivatives thereof and    mixtures thereof.-   48. The composition of any of paragraphs 41-47, wherein the ratio of    curcumin derivative and the compound is from about 99:1 to about    1:99.-   49. The composition of any of paragraphs 41-48, wherein the    composition comprises at least 50% the curcumin derivative-   50. The composition of any of paragraphs 41-49, wherein the    composition comprises a curcumin derivative and aspirin; a curcumin    derivative and a salicylic acid conjugate; a curcumin derivative and    curcumin; aspirin and di(acetylsalicyloyl)-curcumin; aspirin and    monoacetylsalicyloyl-curcumin; aspirin and diacetyl-curcumin;    aspirin and monoacetyl-curcumin; aspirin and diaglutaroyl-curcumin;    aspirin and monoglutaroyl-curcumin; aspirin and    di-gluocose-glutaroyl-curcumin; aspirin and    mono-gluocose-glutaroyl-curcumin; aspirin and    monolinoleol-curcumnin; aspirin and di-linoleoyl-curcumin; a    salicylic acid conjugate and di(acetylsalicyloyl)-curcumin; a    salicylic acid conjugate and monoacetylsalicyloyl-curcumin; a    salicylic acid conjugate and diacetyl-curcumin; a salicylic acid    conjugate and monoacetyl-curcumin; salicylic acid conjugate and    diaglutaroyl-curcumin; salicylic acid conjugate and    monoglutaroyl-curcumin; salicylic acid conjugate and    di-gluocose-glutaroyl-curcumin; a salicylic acid conjugate and    mono-gluocose-glutaroyl-curcumin; salicylic acid conjugate and    monolinoleol-curcumnin; or salicylic acid conjugate and    di-linoleoyl-curcumin.-   51. A pharmaceutical composition comprising a turmeric oil extract    of any of paragraphs 1-10, a curcumin derivative of any of    paragraphs 11-28, or a composition of any of paragraphs 29-50, and a    pharmaceutically acceptable carrier or excipient.-   52. A method of treating inflammation or an inflammatory disease or    condition in a subject, the method comprising: administering a    therapeutically effective amount of a turmeric oil extract of any of    paragraphs 1-10, a curcumin derivative of any of paragraphs 11-29, a    composition of any paragraphs 29-50, a pharmaceutical composition of    paragraph 51, a salicylic acid conjugate, a curcumin ether    derivative to a subject in need thereof, wherein the salicylic acid    conjugate is of formula (II):

-   -   wherein R⁸ is a carbohydrate; R⁹ is H, optionally substituted        alkyl, optionally substituted alkenyl, optionally substituted        alkynyl, or optionally substituted acyl; and analogs,        derivatives, isomers, prodrugs, and pharmaceutically acceptable        salts thereof,    -   and wherein the curcumin ether derivative is of formula (III):

-   -   wherein R₂ and R₄ are both CH₃; and R₁ and R₃ are both alkyl or        one of R₁ and R₃ is H and the other is alkyl.

-   53. The method of paragraph 52, wherein the inflammatory disease or    condition is an inflammatory or allergic disease, an autoimmune    disease, a graft rejection, or another disease in which undesired    inflammatory responses needs to be inhibited.

-   54. The method of any of paragraphs 52-53, further comprising a step    of diagnosing a subject as having an inflammatory disease before    onset of said administration.

-   55. The method of paragraph 52, wherein the subject has been    previously diagnosed with inflammation, inflammatory disease or    condition.

-   56. The method of paragraph 55, the method further comprising a step    of selecting a subject who has inflammation, inflammatory disease or    condition before onset of said administration.

-   57. The method of any of paragraphs 52, wherein the subject is    undergoing treatment for inflammation or inflammatory disease or    condition before onset of said administration.

-   58. The method of any of paragraphs 52-57, wherein the subject is a    mammal

-   59. The method of any of paragraphs 52-58, wherein the subject is    human.

-   60. The method of any of paragraphs 52-59, wherein the    therapeutically effective amount is 1 μg/kg to 150 mg/kg of body    weight of the subject.

-   61. The method of any of paragraphs 52-60, wherein said    administration is daily, every third day, every fourth day, every    fifth day, once-a-week, once-two-weeks, or once-a-month.

-   62. A method of treating cancer or metastasis in a subject, the    method comprising: administering a therapeutically effective amount    of a turmeric oil extract of any of paragraphs 1-10, a curcumin    derivative of any of paragraphs 11-29, a composition of any    paragraphs 29-50, a pharmaceutical composition of paragraph 51, a    salicylic acid conjugate, or a curcumin ether derivative to a    subject in need thereof, wherein the salicylic acid conjugate is of    formula (II):

-   -   wherein R⁸ is a carbohydrate; R⁹ is H, optionally substituted        alkyl, optionally substituted alkenyl, optionally substituted        alkynyl, or optionally substituted acyl; and analogs,        derivatives, isomers, prodrugs, and pharmaceutically acceptable        salts thereof,    -   and wherein the curcumin ether derivative is of formula (III):

-   -   wherein R₂ and R₄ are both CH₃; and R₁ and R₃ are both alkyl or        one of R₁ and R₃ is H and the other is alkyl.

-   63. The method of paragraph 62, wherein the cancer is selected from    the group consisting of adenocarcinoma, lymphoma, blastoma,    melanoma, sarcoma, leukemia, squamous cell cancer, small-cell lung    cancer, non-small cell lung cancer, gastrointestinal cancer,    Hodgkin's and nonHodgkin's lymphoma, pancreatic cancer,    glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder    cancer, breast cancer, colon cancer, colorectal cancer, endometrial    carcinoma, salivary gland carcinoma, kidney cancer, basal cell    carcinoma, melanoma, prostate cancer, vulval cancer, thyroid cancer,    testicular cancer, esophageal cancer, head cancer, and neck cancer.

-   64. The method of any of paragraphs 62-63, further comprising a step    of diagnosing a subject as having cancer before onset of said    administration.

-   65. The method of paragraph 62, wherein the subject has been    previously diagnosed with cancer.

-   66. The method of paragraph 65, the method further comprising a step    of selecting a subject who has cancer before onset of said    administration.

-   67. The method of any of paragraphs 62, wherein the subject is    undergoing treatment for cancer before onset of said administration.

-   68. The method of any of paragraphs 62-67, wherein the subject is a    mammal

-   69. The method of any of paragraphs 62-68, wherein the subject is    human.

-   70. The method of any of paragraphs 62-69, wherein the    therapeutically effective amount is 1 μg/kg to 150 mg/kg of body    weight of the subject.

-   71. The method of any of paragraphs 62-70, wherein said    administration is daily, every third day, every fourth day, every    fifth day, once-a-week, once-two-weeks, or once-a-month.

To the extent not already indicated, it will be understood by those ofordinary skill in the art that any one of the various embodiments hereindescribed and illustrated can be further modified to incorporatefeatures shown in any of the other embodiments disclosed herein.

The following examples illustrate some embodiments and aspects of theinvention. It will be apparent to those skilled in the relevant art thatvarious modifications, additions, substitutions, and the like can beperformed without altering the spirit or scope of the invention, andsuch modifications and variations are encompassed within the scope ofthe invention as defined in the claims which follow. The followingexamples do not in any way limit the invention.

EXAMPLES Example 1 Extraction and Purification of Turmeric Oil

Turmeric Oil Extraction:

Turmeric powder obtained from fresh turmeric roots (95 grams) was takenin a Whatman filter paper pouch, stirred with 1.2 L of hexane 24 hoursand the solvent was concentrated to get a red oil, 5.2 grams. Theresidue in pouch was stirred over night with 500 mL of hexane andconcentrated to get 0.5 g. The combined yield, 5.7 grams.

Turmeric Oil Purification by Successive High Vacuum Distillations:

First Distillation of 5.7 g of crude extract under high vacuum (<1 torr)gave a fraction, 3.54 g at 115 to 135° C., and an un-distilled portion.Second distillation of 3.54 g of distilled product under high vacuum (<1torr) gave 3.10 g of oil. Third distillation gave 2.64 grams at 100-111°C. (<1.00 torr). Fourth distillation gave three fractions: (1) 0.57 g ofmaterial at 100-120° C. (<1.00 torr) (also referred to as NJ-78-11herein); (2) 1.20 g of material at 120-123° C. (<1.00 torr) (alsoreferred to as NJ-78-12 herein); and (3) 0.26 g at 124° C. (<1.00 torr)(also referred to as NJ-78-13 herein). In some embodiments, a turmericoil extract described herein corresponds to the NJ-78-12 fraction.

Column Chromatography Purification of Distilled Turmeric Oil:

In some experiment, fractions NJ-78-11 and NJ-78-13 from differentdistillations were combined together and purified by columnchromatography. In one experiment, ˜8.0 grams of combined NJ-78-12 andNJ-78-13 mixture were purified on a silica gel (150 grams) column andfractions shown in Table 1 were obtained. Based on GC-MS and NMR,NJ-78-12 fraction, consisted of ar-turmerone (53.19%), α-turmerone(38.30%), β-turmerone (8.72%). NJ-78-12: HPLC: (RT, min) 2.39, 2.86,3.62, 4.26; NMR: CDCl₃ (6) 0.93 m, 3H, 1.26-1.32 m, 2.5H, 1.89 s, 3H,1.96 s, 3H, 2.03-2.22 m, 8H, 2.24-2.50 m, 2H, 3.3 m, 0.02H, 4.75 bs, 5.5bs, 5.6-5.75 m, 5.80-5.82 d, 6.04 s, 6.08 s, 6.2 d, 7.12 s. GC-MS, RT:9.79 (216, 132, 119, 105, 91) 10.20 (216, 201, 132, 119, 91, 83), 10.45(218, 120, 105, 91, 83), 10.78 (218, 135, 123, 107, 91, 83)

Fractions MT-133-1 and MT-133-3 showed one single compound on TLC. TheMT-133-1 fractions showed a single compound with an R_(f) of 0.67 by TLC(ethyl acetate/hexane 15/85) and the fraction MT-133-3 showed a singlecompound with an R_(f) of 0.42 by TLC. Accordingly, in some embodiments,a turmeric oil extract of the invention corresponds to fraction MT-133-1and/or MT-133-3.

Based on NMR and GC-MS, MT-133-3 consists of α-turmerone (77.78%),β-turmerone (20.37%) and ar-turmerone (1.85%); TLC (ethylacetate/hexane; 15/85) Rf=0.42; HPLC (RT, min) 3.41; Elemental analysisC, 73.86%; H, 8.79%. NMR: CDCl₃ (6) 0.91, t, J=6.16 Hz) 8H, 1.73 s,1.91, s, 2.21-2.29, 2.55, 4.78 d (J=6.4 Hz), 5.46, s, 5.6-5.8, m, 5.82,d, J=9.7 Hz), 6.07, d, J=1.1 Hz), 6.17-6.20, m, 7.29, s. GC-MS, RT(m/z): 10.20 (218, 119, 105, 83), 10.43 (218, 120, 105, 91, 83)

TABLE 1 Purification of distilled turmeric oil Eluting solvent VolumeFractions Quantity Hexane 1 L MT-133-1 0.14 g MT-133-2 0.04 g 0.5% Ethylacetate, Hexane 1 L — —   1% Ethyl acetate, Hexane 1 L MT-133-3 2.44 gMT-133-4 1.86 g   2% Ethyl acetate, Hexane 2 L MT-133-5 0.82 g MT-133-60.22 g MT-133-7 0.04 g   5% Ethyl acetate, Hexane 1 L MT-133-8 0.10 gMT-133-9 0.06 g Methanol 250 ml  MT-133-10 1.32 g Total obtained fromcolumn 7.04 g

Example 2 Curcumin Derivatives

Curcumin Acetate:

A mixture of curcumin (1.0 g), acetic anhydride (20 ml) and pyridine (2ml) were heated at 70° C. with stirring for 40 minutes and 80° C. for 1hour and at 60° C. over night under dry conditions. The mixture wascooled to room temperature, added to 50 mL of ice coldwater. The mixturewas extracted with chloroform (3×40 mL). The organic layer was stirredwith 5% sodium bicarbonate solution for 0.5 hour. The organic layer wasseparated and washed with 1N HCl (2×50 mL), brine (2×40 mL), dried(anhydrous Na₂SO₄), concentrated under reduced pressure to get yellowsolid, 1.1 g. TLC indicated no curcumin present. Column purification:The product (0.35 g) was purified on 25 grams of silica gel columneluting with 10% ethyl acetate and hexane. Yield, 0.34 g; NMR consistentwith structure.

Glutaric Ester of Curcumin:

Curcumin (1.00 g) was dissolved in tetrahydrofuran (50 ml) in a 250 mLround bottom flask under a flow of nitrogen gas. Dimethyl amino pyridine(56 mg) and triethyl amine (1.33 ml) were added. The color changed fromyellow to light red. Glutaric anhydride (50 ml) was dissolved in 5 mL oftetrahydrofuran and added in drops using a pressure equalizing additionfunnel. The mixture was refluxed and stirred over night. The mixture wascooled and concentrated. 50 mL of ethyl acetate was added, cooled withice, 20 mL of 0.5 N HCl was added. The organic layer was separated andwashed with brine. The brine was combined with the HCl wash, and thecombined solution was extracted with 25 mL of ethyl acetate. Thecombined ethyl acetate layers were dried over anhydrous sodium sulfate,filtered, concentrated under vacuum to get 1.30 g. TLC indicated nocurcumin, NMR consistent with structure.

Curcumin-Acetylsalicylate:

Acetyl salicylic acid (1.0 g) was mixed with thionyl chloride (1.0 mL)and mixture was heated in a 50 mL round bottom flask under nitrogen at80° C. for 1 hour. The excess thionyl chloride was removed bydistillation, dried in high vacuum to get acetyl salicyloyl chloride asa viscous liquid, ˜1.1 g. Used for next step with out furtherpurification. Curcumin (0.50 g) was taken in 10 mL of DMF in a 50 mLround bottom flask under nitrogen and pyridine was added. Acetylsalicyloyl chloride (˜1.1 g) was taken in 5 mL of DMF and was added dropwise from a pressure equalizing separating funnel. The solution turneddarker in color. The mixture was heated at 90° C. for 1 hour and then atroom temperature overnight. To the reaction mixture 100 mL of water wasadded. A solid formed which was filtered, the solid was stirred with 30mL of NaHCO₃ solution. The solid was filtered, washed with water, driedunder high vacuum to yield 1.10 g of the product, curcumin acetylsalicylate or curcumin-aspirin, NMR was consistent with the structure.

Curcumin-glutarate: In a 500 mL round bottom flask curcumin (3.00 g),DMAP (168 mg) and triethyl amine (4 ml) were taken in 150 mL of THF.Glutaric anhydride (2.35 ml) in 15 mL of THF was added drop-wise underanhydrous conditions. The mixture was refluxed for 20 hours at 80° C.The solution was evaporated in a rotovop. The thick residue was taken in150 mL of ethyl acetate, washed with 50 mL of 0.5 N HCl. The HCl washwas extracted with 50 mL of ethyl acetate. The combined ethyl acetateextracts were dried over anhydrous sodium sulfate and was concentratedunder vacuum to get 4.80 g. This was co-evaporated with ether severaltimes to get a solid, 2.5 g. NMR was consistent with the structure.

Curcumin Diacetone Glucose-Glutarate:

A mixture of curcuminglutarate (0.50 g), diacetoneglucose (0.60 g),dicyclohexylcarbodiimide (50 mg), and ethyl acetate (50 mL) was stirred19 hours at 25° C. The solution was filtered, washed with 0.5 N HCl (30mL), 5% NaHCO₃ solution (30 mL), brine (30 mL), dried (anhydrousNa₂SO₄), concentrated under vacuum 1.10 g.

Curcumin Glucose Glutarate:

A mixture of Curcumin Diacetone-glucose glutarate (0.44 g) and aceticanhydride, 70% (25 ml) was refluxed for 2 hours, concentrated, dissolvedin 50% methanol, water, filtered concentrated, 0.16 g. The solid wastreated with 25 mL, 70% acetic acid, heated at 70° C. to 80° C. for 20hours, concentrated and dried in high vacuum, 0.20 g.

Glyceryl Lipid Conjugates of Curcumin:

Curcumin may be conjugated to glyceryl lipids through a linking groupsuch as succinoyl or glutaryl group. Such conjugates would be expectedto facilitate the uptake of the drug through the lipid bilayer of cellmembranes. The mono or di-lipid conjugates can be obtained bycontrolling the molar ratios of curcumin, the linking compounds and theglyceryl lipids. The glyceryl lipids may include glycerol with one ortwo fatty acids attached with ester bonds or fatty alkyl groups withether linkage. Different fatty acids may be used for the glyceryllipids. This can include saturated as well as mono and poly unsaturatedfatty acids. The unsaturated fatty acids can include omega three fattyacids. The following scheme illustrates the synthesis method.

DMAP: Dimethylamino pyridine; TEA: Triethylamine

Curcumin mono or diglutarate can be obtained by controlling theanhydride ratio (one or two equivalents respectively).

The mono- and di-lipid conjugate ratio can be controlled by using one ortwo equivalents of the glyceryl lipids respectively.

Curcumin-Diglutarate-Distearin Monoester Conjugate:

A mixture of curcumin diglutarate (115 mg, 0.18 mmoles), distearin (113mg, 0.18 mmoles), dicyclohexylcarbodiimide (54 mg, 0.18 mmoles),dimethylaminopyridine (14 mg), ethylacetate (10 mL) and dichloromethane(15 mL) was taken in a 50 mL round bottom flask, the solvents wereanhydrous (dried over molecular sieves), and the mixture was stirredovernight. The yellow solution was concentrated and was purified on asilica gel column eluting with ethyl acetate-hexane (0 to 100% ethylacetate). Yield, 119 mg. NMR confirmed the structure.

Example 3 Aspirin-Glucose Conjugate

Aspirin, is a widely used and relatively safe non-steroidalanti-inflammatory drug (NSAID), and it has been used for over hundredyears. The main uses of aspirin can be summarized as an analgesic,antipyretic, anti-rheumatic agent and anti-inflammatory agent, may beused to prevent stroke, to prevent heart attack and to prevent cancer,may be used to treat AIDS and to treat problems associated with diabetes(Fowler). Aspirin is a non-selective cyclooxygenase inhibitor.Cyclooxygenase derived prostaglandins are involved in inflammatoryactivity. Prostaglasndins also are involved in gastrointestinalprotection and vascular homeostasis. Aspirin acetylates Ser530 hydroxylgroup in the COX binding site of COX-1 and COX-2. Aspirin is a muchstronger inhibitor of COX-1 than of COX-2. See for example, Mead, E. A.,Smith, W. L. & DeWitt, d. L., J. Biol Chem., 1993, 268, 6610-6614 andYeomans, N.D., J Gastroenterol Hepatol. 2010 Nov. 10. doi:10.1111/j.1440-1746.2010.06569.x. [Epub ahead of print], content of bothof which is herein incorporated by reference in its entirety. Aspirin isnot very water soluble, only 0.33 g in 100 mL. Some of the undesirableside effects of aspirin results from undissolved particles in thegastrointestinal mucosa casing ulcers and bleeding. The solubility canbe increased considerably by conjugating aspirin to glucose moleculethrough a systematic approach on a specified carbon of glucose.

Synthesis of Glucose-Aspirin:

A glucose-aspirin conjugate was synthesized as shown below.

Hydroxyl protected glucose, 1,2:5,6-diisopropylglucose, was treated withaspirin in presence of dicyclohexylcarbodiimide (DCC) to form the acetylsalicylic acid ester on the fourth carbon of glucose. This ester wastreated with acetic acid to remove the acetonide protecting groups toobtain glucose acetyl salysilate (glucose aspirin).

Synthesis of1,2:5,6-di-O-isopropylidende-3-(2′-acetyloxybenzoyl)-D-glucose

Acetylsalicylic acid (12 g, 0.07 moles) was dissolved in 300 mL of ethylacetate in a 1 liter round bottom flask fitted with anhydrous calciumchloride guard tube. Dicyclohexyl-carbodiimide (8.24 g, 0.04 moles) wasadded to it resulting in the formation of a white precipitate.Diacetone-D-glucose (10 g, 0.04 moles) was added along withdimethylaminopyridine (4.88 g, 0.04 moles). The mixture was stirred for3 hours at room temperature. The solid byproduct, dicyclohexyl urea, wasremoved by filtration. The filtrate was washed with 10% potassiumcarbonate (2×300 mL), 10% hydrochloric acid (2×300 mL) and with brine(300 mL). The ethylacetate solution was dried over anhydrous sodiumsulfate and concentrated to get 17.8 g. The crude product, diacetoneglucose salicylate (compound 3, scheme 1), was purified over silica gelcolumn to get 8.98 g of product. NMR (CDCl₃, δ): 8.0 (dd, 1H), 7.6 (m,1H), 7.33 (m, 1H), 7.13 (dd 1H), 5.9 (m, 1H), 5.4 (d, 1H), 4.61 (d, 1H),4.20-4.37 (m, 2H), 4.03-4.15 (m, 2H), 2.37 (s, 3H), 1.28, 1.32, 1.41,1.62 (our singlets, 12H).

Synthesis of 3-(2′-Acetyloxybenzoyl)-D-glucose (Glucose-3-aspirin)

A mixture of diacetone glucose salicylate (7.6 g) and 70% acetic acid(150 mL) was taken in a 500 mL round bottom flask. The mixture wasrefluxed for 2 hours under argon. The mixture formed a clean solutionand it was concentrated under reduced pressure and dried under highvacuum. Column purification (silica gel, methylene chloride/methanol)yielded the product, glucose-3-aspirin (compound 5) as a white solid,1.3 g. NMR (CDCl₃, 6): 10.6 (s, 1H), 7.9-8.1 9 m, 1H), 7.5 (m, 1H),6.8-7.2 (m, 2H), 5.3 (m, 2H), 4.2-4.6 (m, 2H), 3.6-3.8 (m, 2H), 3.0-3.2(m, 1H), 3.43 (bs, 3H), 1.65 (bs, 3H).

Elemental analysis: C₁₅H₁₈O₉, Calculated: C, 52.63; H, 5.30 Found: C,52.51; H, 5.42. Mass: M+1, m/z 343.0999 (M+H calculated formula:C₁₅H₁₉O₉. Solubility of glucose-3-aspirin in water: 2.32 g/100 mL,Aspirin solubility: 0.33 g/100 mL (1 g in 300 mL water at 25° C.)(MerckIndex, pp. 134, vol. 11, 1989). Thus, the aspirin-glucose conjugate was700 times more soluble than aspirin.

Example 4 In Vitro Enzyme Hydrolysis of Glucose Aspirin Conjugate

A 25% solution of human serum in 0.01M phosphate buffer was taken andthe test compound (glucose aspirin or aspirin) were added to get aconcentration of 0.2 mg/mL. The hydrolysis for the rate of release ofsalicylic acid and the rate of decomposition of the test compound weremeasured by HPLC. As shown in Tables 2 and 3, glucose aspirin hydrolysedat a much slower rate than aspirin in human serum.

TABLE 2 In vitro enzyme hydrolysis of salicylic acid vs glucose- aspirinconjugate Salicyclic acid Glucose-aspirin Time Concentra- RateConcentra- Rate (min) tion (M) (M/min) tion (M) (M/min) 0 0 0.00E+001.98E−04   0.00E+00 30 0.00E+00 0.00E+00 2.03E−04   1.91E−07 60 2.70E−06 4.50E−0.8 1.84E−04 −2.24E−07 90 2.75E−06  3.06E−0.8 1.65E−04 −3.62E−07120 1.09E−05 9.11E−07 1.62E−04 −2.96E−07 180 3.54E−05 1.97E−07 1.19E−04−4.38E−07 Average rate: 9.08E−08 −3.30E−07 Std.dev. 7.51E−08   2.46E−07

TABLE 3 In vitro enzyme hydrolysis of salicylic acid vs acetylsalicyclicacid Salicyclic acid Acetylsalicyclic acid Time Concentration RateConcentration Rate (min) (M) (M/min) (M) (M/min) 0 0 0.00E+00 4.57E−04  0.00E+00 30 1.95E−05 6.49E−07 4.15E−04 −1.40E−06 60 3.39E−05 5.65E−073.99E−04 −9.81E−07 90 4.98E−05 5.53E−07 2.48E−04 −2.32E−06 120 7.49E−056.24E−07 2.70E−04 −1.56E−06 180 1.07E−04 5.94E−07 1.74E−04 −1.58E−06Average rate: 5.97E−07 −1.57E−06 Std.dev. 3.17E−08   4.85E−07

Example 5 Animal Studies with Glucose Aspirin Conjugates

Materials and Methods:

Animals were kept under 13 hour day night cycle Animals were givenstandard diet and water was provided ad, libitum. All animals wereacclimatized for at least one week before the experimental session.Animals were divided in to five groups. Each group consisted of 8animals. Group 1 would get standard drug, aspirin at 100 mg/kg and group2 was given aspirin 200 mg/kg i.p. Glucosyl acetyl salicylate was usedin two dose levels. Group 3 was given the glucosyl acetyl salicylate at100 mg/kg and group 4 was given this drug at 200 mg/kg i.p.

Carrageenan Induced Rat Paw Edema:

Pedal inflammation in albino rats of either sex was produced accordingto the method of Winter et al., (Proc Soc Exp Biol Med 1963; 111:544-7). An injection was made with 0.1 ml of 1% carrageenan (SIGMA)suspension into the right hind foot of each rat in the plantar region.Groups 3 and 4 rats were treated intraperitoneally (i.p.) withglucosylacetyl salicylate 30 minutes before carrageenan injection.Control group 5 was given 0.5 ml saline and groups 1 and 2 receivedaspirin. Increase in linear paw circumference was measured by tying apiece of cotton thread round the rat's paw, and noting the point ofintersection of two ends on a meter scale. This was taken as an index ofincrease in paw volume, which is a measure of oedema (Bamgbose S O A,Noamesi B K. Planta Med 1981; 42: 392-6). Measurements were takenimmediately before and at 1, 2 and 3 hours after carrageenan injection.The inhibitory activity was calculated according to the followingformula (Awe S O, Olajide O A, Adeboye J O, Makinde J M. Indian JPharmacol 1998; 30: 38-42):

${{Percent}\mspace{14mu} {inhibition}} = \frac{{\left( {{Ct}\text{-}{Co}} \right)\mspace{14mu} {control}} - {\left( {{Ct}\text{-}{Co}} \right)\mspace{14mu} {treated} \times 100}}{\left( {{Ct}\text{-}{Co}} \right)\mspace{14mu} {control}}$

Ct-Linear paw circumference 3 hours after carrageenan injection andCo-Linear paw circumference before carrageenan injection.

Tail flick test: Analgesic activity was measured by the tail flickmethod, using the analgesiometer (Techno, Lucknow, India) as describedby D'Armour and Smith (D'Armour F E, Smith D L. A method for determiningloss of pain sensation. J Pharmacol Exp Ther 1941; 72: 74-9). Rats werescreened for tail flip reaction with a cut-off time of 5 seconds. Foreach animal, the tail flick latency was obtained thrice before drugadministration and mean was used as pre-drug latency. The tail flicklatencies were measured at 0, 0.5, 1, 1.5, 2 and 3 h afteradministration chemicals. For animals that did not respond within thecut-off time of 10 seconds, the value of the cut-off time was consideredas latency period for that animal (Ramabadran K, Bansinath M. A criticalanalysis of the experimental evaluation of nociceptive reactions inanimals. Pharmaceutical Res 1986; 3: 263-70). Results was expressed as %maximum possible effect (MPE) (Bishnoi M, Patil C S, Kumar A, Kulkarni SK. Analgesic activity of acetyl-11-keto-beta-boswellic acid, a5-lipoxygenase-enzyme inhibitor. Indian J Pharmacol 2005; 37: 255-56):

${\% \mspace{14mu} M\; P\; E} = {\frac{\left( {{{post}\mspace{14mu} {treatment}\mspace{14mu} {latency}} - {{pretreatment}\mspace{14mu} {latency}}} \right)}{{Pretreatment}\mspace{14mu} {latency}} \times 100}$

Statistical analysis: Data was analyzed using analysis of variance(ANOVA), t-test, Mann-Whitney and Chi-square tests. P<0.05 wasconsidered as statistically significant.

FIG. 5 shows the effect of drugs on tail flick latency in all groups.There was significant increase in the tail flick latency with thestandard drug aspirin 100 mg/kg (Group1) and aspirin 200 mg/kg (Group 2)at all times intervals except 180 minutes as compared to control group(p<0.05). The increase was there till 120 minutes. There was nostatistical significant change in the tail flick latency in the controlgroup (Group 5). In test group there was significant increase in tailflick latency with both dose levels i.e. 100 mg/kg (Group 3) and 200mg/kg (Group 4). The increase was there till 120 minutes in group 3 and90 minutes in group 4. % MPE was more with 100 mg/kg as compared to 200mg/kg.

Aspirin 100 mg/kg and 200 mg/kg significantly increased latency time ascompared to control group (p<0.05) at 60 minutes. The latency time wassignificantly more at 60 minutes with both doses of aspirin as comparedto 100 mg/kg dose of test compound (FIG. 5). Percentage (%) MPE was morewith 100 mg/kg as compared to 200 mg/kg (FIG. 6). Percentage MPE was atpeak at 120 minutes in all the groups. There was no statisticallysignificant difference in the latency time with test compound ascompared to control group.

These results demonstrate that Test compound in doses of 100 and 200mg/kg significantly suppressed carrageenan-induced paw edema in rats anddemonstrated significant analgesic activity in tail flick models.

Example 6 Animal Studies with Turmeric Oil Extracts

Anti-Inflammation and Analgesic Studies:

Albino rats obtained from central animal house of institute were used inthe study. The animals were kept under 12 hour day night cycle Animalswere given standard diet and water was provided ad libitum. All animalswere acclimatized for at least one week before the experimental sessionAnimals were divided into twelve groups in the carrageenan induced pawedema and tail flick method. Each group consisted of 4 animal each. Thegroups received various chemicals as given in Table 4.

TABLE 4 Chemicals and dosage given to the various animal groups Group #Chemical Given^(a) Dos/volume 0 DMSO 0.3 ml 1 NJ1 50 mg/kg 2 NJ1 100mg/kg 3 NJ1 200 mg/kg 4 NJ2 50 mg/kg 5 NJ2 100 mg/kg 6 NJ2 200 mg/kg 7NJ3 50 mg/kg 8 NJ3 100 mg/kg 9 NJ3 200 mg/kg 10 Aspirin 100 mg/kg 11Aspirin 200 mg/kg 12 Normal Saline 0.3 ml ^(a)NJ1 = turmeric oildistillate 115° C.-120° C./high vacuum (~100 torr), NJ2 = Omega-3,DHA/EPA—fish oil, and NJ3 = NJ1 + NJ2 (1:1)

Carrageen Induced Paw Edema:

There was significant increase in paw measurements (mm) after carrageeninjection in all the groups (Table 5). The increase was maximum in group0 and 12 (control groups) verifying the effectiveness of the method. In% inhibition of paw edema, all drugs in various groups inhibited theedema. The standard used (aspirin) significantly inhibited thedevelopment of the edema as compared to control groups confirming thevalidity of the method (FIG. 9). The formulation NJ1 (50 mg/kg)significantly inhibited edema at 1 hour as compared to control groups(FIG. 10). This formulation also inhibited edema at 2 and 3 hours ascompared to group 1 (DMSO), but the inhibition was not significant ascompared to aspirin (group 5 and 6). Group 3 (NJ1 100 mg/kg)significantly inhibited edema as compared to DMSO (group 1) at 1 houronly (FIG. 11). Group 3 (2 and 3 hour) and group 4 (all times) inhibitededema less than the standard drug aspirin (FIG. 12). NJ2 and NJ3 did notsignificantly inhibit edema as compared to control and aspirin. In factthe percentage inhibition was significantly less with NJ2 (100 and 200mg/kg-group 3 and 4) and NJ3 (200 mg/kg-group 4). The % inhibition ofpaw edema at 3 hours was maximum for NJ 1 (50 mg/kg) followed by aspirin200 mg/kg and aspirin 100 mg/kg.

TABLE 5 Paw circumference (mm) in various groups Groups 0 h 1 h 2 h 3 h0 2.75 ± 0.3  3.475 ± 0.19*  3.55 ± 0.42* 3.475 ± 0.33* 1 2.775 ± 0.13 3.075 ± 0.10*  3.1 ± 0.43 3.225 ± 0.21* 2 2.65 ± 0.24 2.825 ± 0.26*3.125 ± 0.26* 3.325 ± 0.24* 3 2.75 ± 0.10 3.025 ± 0.13*  3.35 ± 0.19*3.525 ± 0.10* 4 3.25 ± 0.21 3.35 ± 0.30 3.375 ± 0.19* 3.425 ± 0.29  5 3.4 ± 0.12  3.9 ± 0.73*  3.9 ± 0.26* 3.775 ± 0.22* 6 2.725 ± 0.30 3.575 ± 0.17* 3.25 ± 0.26  3.55 ± 0.37* 7 3.275 ± 0.19   3.55 ± 0.10* 3.5 ± 0.35* 3.525 ± 0.38  8 3.025 ± 0.13   3.35 ± 0.19* 3.25 ± 0.37 3.25 ± 0.38* 9 2.675 ± 0.22   3.65 ± 0.12* 3.125 ± 0.51* 3.525 ± 0.23*10 3.125 ± 0.13  3.475 ± 0.15   3.6 ± 0.14* 3.475 ± 0.10* 11  3.3 ± 0.183.425 ± 0.21  3.575 ± 0.13*  3.55 ± 0.10* 12 3.525 ± 0.53   4.15 ± 0.24* 4.4 ± 0.22*  4.55 ± 0.24* *p < 0.05 as compared to 0 hour

Tail Flick:

The increase no increase in tail flick latency in group 0 and 12(control groups) verifying the usefulness of the method (Table 6). Therewas significant increase in tail flick latency (seconds) with somedrugs. The % MPE was significantly more with aspirin as compared tocontrol groups (FIG. 13) confirming the validity of the method (FIG. 5).The % MPE was significantly more with formulation NJ1 (50 mg/kg) at 180minutes as compared to DMSO (FIG. 14). The % MPE was less with NJ2 andNJ3 as compared to aspirin at all dose levels and at all times (FIG.15). The % MPE with NJ3 was less as compared to aspirin (FIG. 16).

TABLE 6 Tail flick latency time (seconds) in various groups Groups 0 min30 min 60 min 90 min 120 min 180 min 0 2.02 ± 0.26 2.37 ± 0.39 2.13 ±0.38 2.24 ± 0.33 2.10 ± 0.70 1.915 ± 0.13  1 1.33 ± 0.24 1.46 ± 0.43 2.26 ± 0.38*  1.50 ± 0.20* 1.39 ± 0.18 1.105 ± 0.13  2 1.295 ± 0.22 1.39 ± 0.21  2.50 ± 0.13* 1.38 ± 0.24 1.55 ± 0.37 1.125 ± 0.25  3 1.65 ±0.27  2.27 ± 0.54*  2.51 ± 0.39*  3.47 ± 0.43*  2.76 ± 0.75* 1.85 ± 0.304 1.71 ± 0.54 2.40 ± 0.58 2.36 ± 0.15 2.105 ± 0.15  1.96 ± 0.10 2.22 ±0.23 5 1.79 ± 0.22 1.97 ± 0.33 1.83 ± 0.13 2.01 ± 0.70 1.855 ± 0.26 1.76 ± 0.14 6 1.76 ± 0.30 1.80 ± 0.29 1.97 ± 0.70 1.79 ± 0.69 1.78 ±0.17 1.98 ± 0.31 7 1.63 ± 0.22 1.21 ± 0.20 1.68 ± 0.18 1.67 ± 0.71 1.45± 0.17 1.29 ± 0.49 8 1.77 ± 0.47 1.63 ± 0.51 2.00 ± 0.29 1.66 ± 0.531.61 ± 0.53 1.43 ± 0.29 9 2.29 ± 0.72 2.31 ± 0.46 1.82 ± 0.53 2.65 ±0.93 1.89 ± 0.57 1.89 ± 0.16 10 1.64 ± 0.27  3.28 ± 0.26*  3.28 ± 0.57* 2.88 ± 0.26*  2.73 ± 0.48* 2.05 ± 0.76 11 2.32 ± 0.46  3.46 ± 0.77* 3.46 ± 0.17* 3.03 ± 0.42 3.12 ± 0.30 2.53.45 12 2.36 ± 0.38 2.377 ±0.17  2.37 ± 0.39 2.28 ± 0.18 1.90 ± 0.24 2.41 ± 0.34 *p < 0.05 ascompared to 0 minutes

Carrageenan-induced hind paw edema is the standard experimental model ofacute inflammation. Carrageenan is the agent of choice for testinganti-inflammatory drugs as it is not known to be antigenic and is devoidof apparent systemic effects. Moreover, the experimental model exhibitsa high degree of reproducibility. Carrageenan-induced edema is abiphasic response. The first phase is mediated through the release ofhistamine, serotonin and kinins whereas the second phase is related tothe release of prostaglandin and slow reacting substances which peak at3 hour. See for example, Vinegar, R., Schreiber, W. & Hugo R., J.Pharmacol. Exp. Ther., 1969, 166: 96-103.

The increase in the paw volume following carrageenan administration inthe control and aspirin treated group corresponds with the findings ofprevious workers. NJ1 produced significant inhibition ofcarrageenan-induced paw edema. The effect produced is equivalent to theeffect of aspirin indicating similar efficacy. The effect with NJ1 wasmore at lower dose. However NJ1 showed dose dependant analgesic activityon tail flick test, higher dose showing more effect. NJ2 and NJ3 did notshow any significant activity, infact their effect was lower than theaspirin. Thus, these results demonstrated that NJ1 has significantanti-inflammatory and analgesic activity in animal models.

Example 7 Anti-Cancer Activity of Turmeric Oil Distillation Fraction andCurcumin Analogs

Assays were performed with 3 cancer cell lines: breast (SKBR3),pancreatic (Panc1), prostate (PC-3) and 1 normal (nor-cancer) cell line(WI-38). Compounds were delivered to the cells after dissolving in DMSO.Five dilutions per compound were tested on a total of 4 cell lines in 96well plates. Cellular inhibition was measured using Alamar Blue. Afteraddition of compound a 24 hour and 48 hour time point using fluorescentplate reader was taken. Data from plate reader was analyzed in thefollowing way: % inhibition bar graphs, % inhibition line graphs andIC50 growth inhibition values. The results of the experiments are shownin FIGS. 17A-25B. IC₅₀ values are summarized in Tables 7-14. In Tables7-14, IC₅₀ units are Molar (M).

TABLE 7 Summary of IC₅₀ values for NJ-58-1, MT-76-1 and curcumin (24hours) Compound Cell line IC₅₀ NJ-58-1 24 SKBR3 1.71E−04 NJ-58-1 24 WI384.39E−03 NJ-58-1 24 PANC1 2.68E−04 NJ-58-1 24 PC3 9.61E−05 MT-76-1 24SKBR3 1.69E−04 MT-76-1 24 WI38 2.98E−04 MT-76-1 24 PANC1 3.54E−05MT-76-1 24 PC3 1.27E−04 Curcumin 24 SKBR3 8.41E−05 Curcumin 24 W1381.00E−03 Curcumin 24 PANC1 4.15E−05 Curcumin 24 PC3 2.95E−05

TABLE 8 Summary of IC₅₀ values for NJ-78-12 and MT-133-1 (24 hours)Compound Cell line IC₅₀ NJ-78-12 SKBR3 5.77E−03 NJ-78-12 WI38 1.40E+03NJ-78-12 PANC1 2.95E−03 NJ-78-12 PC3 1.36E−03 MT-133-3 SKBR3 3.43E−04MT-133-3 WI38 3.24E−04 MT-133-3 PANC1 3.95E−04 MT-133-3 PC3 3.27E−04

TABLE 9 Summary of IC₅₀ values for PMN 11-168 and Aspirin (24 hours)Compound Cell line IC₅₀ PMN II-168 SKBR3 2.01E−04 PMN II-168 WI383.76E−04 PMN II-168 PANC1 3.99E−04 PMN II-168 PC3 8.32E−05 ASPIRIN SKBR37.31E−04 ASPIRIN WI38 6.19E−04 ASPIRIN PANC1 9.75E−04 ASPIRIN PC37.71E−04

TABLE 10 Summary of IC₅₀ values for NJ-81-4 and Taxol (24 hours)Compound Cell line IC₅₀ NJ-81-4 SKBR3 5.54E−07 NJ-81-4 WI38 4.13E−07NJ-81-4 PANC1 9.90E−08 NJ-81-4 PC3 6.48E−08 Taxol 24 SKBR3 6.94E−07Taxol 24 WI38 1.91E−05 Taxol 24 PANC1 1.53E−07 Taxol 24 PC3 5.81E−08

TABLE 11 Summary of IC₅₀ values for NJ-58-1, MT-76-1 and curcumin (48hours) NJ-58-1 48 SKBR3 1.25E−04 NJ-58-1 48 WI38 1.55E−04 NJ-58-1 48PANC1 1.33E−04 NJ-58-1 48 PC3 6.70E−05 MT-76-1 48 SKBR3 1.18E−04 MT-76-148 WI38 1.66E−04 MT-76-1 48 PANC1 1.23E−04 MT-76-1 48 PC3 7.16E−05Curcumin 48 SKBR3 9.94E−05 Curcumin 48 WI38 8.99E−05 Curcumin 48 PANC13.82E−05 Curcumin 48 PC3 2.42E−05

TABLE 12 Summary of IC₅₀ values for NJ-78-12 and MT-133-1 (48 hours)Compound Cell line IC₅₀ NJ-78-12 SKBR3 2.33E−04 NJ-78-12 WI38 2.98E−03NJ-78-12 PANC1 3.22E−04 NJ-78-12 PC3 4.98E−04 MT-133-3 SKBR3 3.18E−04MT-133-3 WI38 3.28E−04 MT-133-3 PANC1 4.02E−04 MT-133-3 PC3 3.34E−04

TABLE 13 Summary of IC₅₀ values for PMN 11-168 and Aspirin (48 hours)Compound Cell line IC₅₀ PMN II-168 SKBR3 8.78E−05 PMN II-168 WI383.10E−04 PMN II-168 PANC1 9.05E−05 PMN II-168 PC3 8.14E−05 ASPIRIN SKBR36.93E−04 ASPIRIN WI38 5.01E−04 ASPIRIN PANC1 7.45E−04 ASPIRIN PC37.82E−04

TABLE 14 Summary of IC₅₀ values for NJ-81-4 and Taxol (48 hours)Compound Cell line IC₅₀ NJ-81-4 SKBR3 3.09E−07 NJ-81-4 WI38 8.12E−04NJ-81-4 PANC1 4.60E−09 NJ-81-4 PC3 1.76E−08 Taxol 48 SKBR3 3.23E−06Taxol 48 WI38 8.00E−05 Taxol 48 PANC1 1.80E−07 Taxol 48 PC3 8.03E−08

Glucose Aspirin (PMN 11-168) vs. Aspirin:

Examining the bar graphs at 24 hours, showed that PMN-II-168 had amodest 20% inhibition of cancer cell growth for the four lowerconcentrations while aspirin did not seem to inhibit growth by anysignificant means at 24 hours. IC₅₀ for PMN-II-168 were found to be2.01E-04, 3.76E-04, 3.99E-04 and 8.32E-05 for SKBR3, WI38 PANC1 and PC-3cells respectively. IC₅₀ for aspirin were found to be 7.31E-04,6.19E-04, 9.75E-04 and 7.71E-04 for SKBR3, WI38 PANC1 and PC-3 cellsrespectively. IC₅₀ values are in Molar (M).

PMN-II-168 at 300 uM did inhibit cancer cell growth very well (up to 60%inhibition) while not playing a significant inhibitory role with thenormal WI-38 cell lines. This was consistent at the variousconcentrations. Aspirin however did not have the same selectivity asPMN-II-168 as WI-38 cells as they were the most efficiently growthrepressed at 24 hours. At 48 hours, while there was little change inaspirin inhibition (except for the highest 300 uM concentration),PMN-II-168 did significantly better. PMN-II-168 inhibited breast cancerline (skbr3) proliferation most efficiently (approximately 90% and 60%inhibition at the 300 uM and 150 uM concentration respectively). It alsoinhibited Panc-1 (˜70% and 60%) and PC-3 (˜85% and 65%) for the twohighest concentrations, 300 uM and 150 uM respectively. Even at thelower concentrations, PMN-II-168 performed significantly better thanaspirin.

Paclitaxel (Taxol) (1 mg) vs. NJ-81-4 (Mixture of Turmeric OilDistillation Fraction NJ-78-12 (10 mg) and Paclitaxel (1 mg)) at 48Hours:

Taxol at the 48 hour treatment point was effective in inhibitingprostate (PC-3) cancer cells at all concentrations (˜60 inhibition) andat the 300 and 150 nM concentrations inhibited the other cancer celllines more modestly. On the other hand, at the same doses, NJ-81-4 wasmore effective as it inhibited Panc-1 and PC-3 by approximately ˜60% atall concentrations when compared to untreated cells. SKBR3 cell growthproliferation was also inhibited with NJ-81-4 albeit more modestly up to(45% inhibition). In addition to the better inhibition, NJ-81-4 did notaffect normal cell growth (WI-38). Taxol however did have some effect onnormal cell growth.

Turmeric Oil Fraction NJ-78-12 and MT-133-3:

At the 48 hour time point bar graphs, NJ-78-12 at the higher doseinhibited the cancer cell lines, but at none of the doses affected thenormal tissue WI-38 cell line. Thus, this compound is specific and lacksa general toxicity to cells or tissue. MT-133-3 behaved similar toNJ-78-12 except that it inhibited WI-38 growth. NJ-81-4 also did verynicely giving good inhibition of cancer cells but not the WI-38 cells.

Curcumin Derivatives Diacetyl Curcumin (NJ-58-1) and Curcumin-Glutarate(MT-76-1):

The curcumin levels of inhibition were generally higher than NJ-58-1 andMT-76-1. An exception was with skbr3 breast cancer cells at 300 uM. Allthree compounds gave a similar response at 24 hours with skbr3. At 24hours, NJ-58-1 was more specific to cancer cells than curcumin. The IC₅₀data for 24 and 48 hour time points is shown in Tables 8 and 12. At 24hours, NJ-58-1 did very well in inhibiting PANC-1 and PC-3 at dosesgreater than 50 uM. It inhibited skbr3 cancer lines at greater than 150uM doses. MT-76-1 at 24 hours behaved similarly except that at the lowerdoses it more specifically inhibited the PANC-1 cell line. Bothcompounds at 24 hours didn't have a very significant affect on the WI-38cell line, which is a normal cell line, i.e. not a cancerous cell line.At 24 hours with NJ-58-1, normal lung cells (WI-38) actually grew orwere inhibited less than 10%, while with curcumin levels of inhibitionat 100, 150 and 300 uM were just over 20%. Accordingly, these resultsdemonstrate that NJ-58-1 is more specific than MT-76-1 and curcuminbecause normal cells behaved similar to cancer cells treated withcurcumin.

Example 8 Evaluation of Anti-Cancer Activity of Compounds

The following compounds and combinations were used in this experiment:Gemcitabine.HCl (NJ-92-1), MT-133-3 (NJ-92-2), Gemcitabine.HCl (2mg)+MT-133-3 (20 mg) (NJ92-3), Paclitaxel (NJ-92-4) and Paclitaxel 2mg+MT-133-3 (20 mg) (NJ-92-5).

Determine LD50 (Lethal Dose, 50%) of Testing Compounds:

Cytotoxicity of the compounds on three different cancer cell lines andone normal cell line was tested. Four human cell lines including bothcancer cell lines (SK-BR-3, PANC-1, PC3) and normal cell line (WI38)were initially employed. The cells (20,000 cells/well) were plated ontoa 96-well plate for 24 hours before treatment. The compounds werediluted in the medium and added to the cells at different concentrationsfor additional 24 hours from 1 ng/ml to 10 ug/ml. MTT assay was thenperformed.

LD50 was not observed for all these compounds under testingconcentrations, demonstrating that these compounds do not havecytotoxicity under the testing conditions (FIGS. 26A-26D).

Determination of Inhibitory Effect of Compounds on Cell Proliferation:

The compounds were tested for cancer cell proliferation inhibition.Based on the data derived from in vitro cytotoxicity, evaluation of cellproliferation was under taken. Four cell lines were plated onto a 96well plate for 24 hours before adding the compounds. Two different doses(20 ng/ml and 2,000 ng/ml) were employed up to 8 days followed by MTTassay. DMSO and Taxol were used as negative control and positivecontrols. Results are shown in FIGS. 27A-27D.

Compound #5 (NJ-92-5) displayed inhibitory effect on PANC-1 cellproliferation after 48 h treatment; the compound #5 inhibited PC3 cellproliferation at early treatment, similar to Taxol; the compound #5showed cytotoxicity on SK-BR-3 cells. No significant difference ofCompound #5 was noted on normal WI38 cells.

Compound #3 (NJ-92-3) had minimal effect on cancer cell proliferation.Significant effect occurred only under high concentration (2,000 ng/ml).

These results indicate that the mechanism of action of compound #5(NJ-92-4) is through inhibition of cancer cell proliferation, but notcytotoxiticy.

Determination of GI₅₀ (50% Inhibition of Cell Growth):

Since the compounds displayed inhibitory effect on cell proliferation, aGI₅₀ experiment was performed. The cells were plated onto a 96 wellplate and treated with different concentrations of the compounds from 1ng/ml to 10 ug/ml for 72 hours followed by MTT assay. The results areshown in FIGS. 28A-28D and GI₅₀ values summarized in Table 15.

TABLE 15 GI₅₀ values of compounds for various cell lines Cell lineNJ-92-1 NJ-92-2 NJ-92-3 NJ-92-4 NJ-92-5 PANC-1 7180 ng/ml 5641 ng/ml6411 ng/ml  1.8 ng/ml  0.6 ng/ml PC3 3847 ng/ml 4616 ng/ml NA  258 ng/ml 6923 ng/ml SK-BR-3 NA 9743 ng/ml 9999 ng/ml 1.15 ng/ml  0.25 ng/ml WI38NA 5385 ng/ml 6411 ng/ml  514 ng/ml  2052 ng/ml

Example 8 In Vivo Effect of Curcumin, Turmeric Oil and Fish OilCombinations

Materials and Methods: The study protocol was approved by IAEC. Albinorats obtained from the central animal house of the institute were usedin the study. The animals were kept under 12 hour day night cycle.Animals were given standard diet and water was provided ad libitum. Allanimals were acclimatized for at least one week before the experimentalsession.

Animals were divided into eight groups in the carrageenan induced pawedema and tail flick method. Each group consisted of 6 animal each. Thegroups received various chemicals as given in Table 16 by oraladministration with food. The following methods were used to evaluatethe activity of the compositions:

Carrageenan Induced Rat Paw Edema:

Pedal inflammation in albino rats of either sex was produced accordingto the method of Winter et al. (Proc. Soc. Exp. Biol. Med., 1963, 111:544-547). An injection was made with 0.1 ml of 1% carrageenan (SIGMA)suspension into the right hind foot of each rat in the plantar region.The various chemicals were given 30 minutes before carrageenaninjection. Increase in linear paw circumference was measured by tying apiece of cotton thread round the rat's paw, and noting the point ofintersection of two ends on a meter scale. This was taken as an index ofincrease in paw volume, which is a measure of edema (Bamgbose, S. O. A.& Noamesi, B. K., Planta. Med., 1981, 42: 392-396). Measurements weretaken immediately before and at 1, 2 and 3 hours after carrageenaninjection. The inhibitory activity was calculated according to thefollowing formula (Awe S O, Olajide O A, Adeboye J O, Makinde J M.Indian J. Pharmacol., 1998; 30: 38-42):

${{Percent}\mspace{14mu} {inhibition}} = \frac{{\left( {{Ct}\text{-}{Co}} \right)\mspace{14mu} {control}} - {\left( {{Ct}\text{-}{Co}} \right)\mspace{14mu} {treated} \times 100}}{\left( {{Ct}\text{-}{Co}} \right)\mspace{14mu} {control}}$

Ct-Linear paw circumference 3 hours after carrageenan injection andCo-Linear paw circumference before carrageenan injection.

Tail flick test: Analgesic activity was measured by the tail flickmethod, using the analgesiometer (Techno, Lucknow, India) as describedby D'Armour and Smith (J. Pharmacol. Exp. Ther., 1941, 72: 74-79). Ratswere screened for tail flip reaction with a cut-off time of 5 seconds.For each animal, the tail flick latency was obtained thrice before drugadministration and mean was used as pre-drug latency. The tail flicklatencies were measured at 0, 0.5, 1, 1.5, 2 and 3 h afteradministration of chemicals. For animals that did not respond within thecut-off time of 10 seconds, the value of the cut-off time was consideredas latency period for that animal (Ramabadran, K. & Bansinath, M.Pharmaceutical. Res., 1986, 3: 263-270).

Results are expressed as % maximum possible effect (MPE) (Bishnoi, etal., Indian J. Pharmacol., 2005, 37: 255-256)

${\% \mspace{14mu} M\; P\; E} = {\frac{\left( {{{post}\mspace{14mu} {treatment}\mspace{14mu} {latency}} - {{pretreatment}\mspace{14mu} {latency}}} \right)}{{Pretreatment}\mspace{14mu} {latency}} \times 100}$

Statistical Analysis:

Data were analyzed using analysis of variance (ANOVA) and t-test. P<0.05was considered as statistically significant.

TABLE 16 Groups Drug Dose/volume 1 Normal saline 0.3 ml 2 Curcumin 100mg/kg 3 Curcumin + fish oil (1:1) 100 mg/kg 4 Curcumin + turmeric oil,BR-132-4 (1:1) 100 mg/kg 5 Curcumin + fish oil + lecithin (4:2.5:1) 100mg/kg 6 Turmeric oil fraction, BR-132-4 100 mg/kg 7 Aspirin 100 mg/kg 8Sod. Bicard. 0.3 ml

Results for paw edema inhibition are shown in Table 17 and FIG. 30. Forpaw edema inhibitory activity curcumin and turmeric oil mixture (group4) at 1 and 3 hours exhibited the maximum activity at a dose of 50 mg/kgfor each of the compounds and this was higher than that of curcumin(group 2) and turmeric oil (group 6) alone at 100 mg/kg doses. At 2hours a combination of curcumin, fish oil and lecithin (group 5) inwhich curcumin is ˜53 mg/kg, had stronger activity than that of curcuminalone at 100 mg/kg. At all time points the combination of curcumin andfish oil (group 3) in which curcumin was at 50 mg/kg, had similar orslightly stronger activity than curcumin at 100 mg/kg.

TABLE 17 Paw circumference (mm) in various groups Groups 0 h 1 h 2 h 3 h1 3.17 3.77 3.45 3.5 2 3.2 3.68 3.45 3.53 3 3.37 3.53 3.68 3.39 4 3.173.7 3.38 3.73 5 3.17 3.57 3.5 3.65 6 2.47 3.43 3.35 3.33 7 3.4 3.82 3.583.67 8 2.47 3.43 3.35 3.33 *p < 0.05 as compared to 0 hour-in allsignificant except at 1 hour in group 4

Results for tail flick latency are shown in Table 18 and FIG. 31. Fortail flick experiment curcumin at 100 mg/kg had stronger activity at 60and 90 minutes compared to the other drugs while the combination ofcurcumin and turmeric oil fraction at 50 mg/kg each had higher activitythan turmeric oil or aspirin alone at 100 mg/kg at 90 minutes.

TABLE 18 Tail flick latency time (seconds) in various groups (*p < 0.05as compared to 0 minutes) Groups 0 min 30 min 60 min 90 min 120 min 180min 1 2.19 2.4 3.07 3.23* 2.47* 2.68 2 2.67 2.58 2.8* 3.18* 3.03 2.68 32.17 2.5 2.4 2.68 2.15 2.18 4 2.48 2.87 2.5 2.77 2.57 2.83 5 2.48 2.382.75 2.07 2.38 2.77 6 1.75 1.9 1.88 1.58 1.87 1.93 7 2.37 2.75* 2.682.57 2.23 2.37 8 2.87 2.67 2.97 2.88 3.05 2.82

These data show that the combination of curcumin and fish oil at 50mg/kg each had strong anti inflammatory activity while curcumin andturmeric oil combination acted as analgesic at 50 mg/kg under theexperimental conditions used.

Example 9 Comparison of Anti-Inflammatory and Analgesic Activities ofFish Oil, Turmeric Oil, Curcumin Diacetate, Curcumin Diglutarate andAspirin

Fish oil, a distillation fraction of turmeric oil and two derivatives ofcurcumin (NJ-104-1 to NJ-104-5) were used in this study to determine theanti-inflammatory and analgesic activities of these drugs by oraladministration and these were compared to aspirin. Anti-inflammatory wasevaluated by carrageenan induced paw edema and analgesic activity bytail flick method in rats.

Albino rats obtained from the central animal house of the institute wereused in the study. The animals were kept under 12 hour day night cycleAnimals were given standard diet and water was provided ad libium. Allanimals were acclimatized for at least one week before the experimentalsession.

Animals were divided into six groups in the carrageenan induced pawedema and tail flick method. Each group consisted of 8 animals each. Thegroups received various drugs orally mixed with food, as given in Table19. Carrageenan-induced rat paw edema and Tail flick test were performedas discussed above in Example 8. Data were analyzed using analysis ofvariance (ANOVA) with Posthoc Bonferroni multi-comparisons. P<0.05 wasconsidered as statistically significant.

TABLE 19 Drugs given to various groups Group Drug Dose/Volume 1 NormalSaline 0.3 mL 2 NJ- 104-1 (Fish oil) 100 mg/kg 3 NJ-104-2 (Turmeric oilfraction, NJ-100-9) 100 mg/kg 4 NJ-104-3 (Fish oil + turmeric oilfraction NJ-100-9) 100 mg/kg (1:1) 5 NJ-104-4 (Curcumin diacetate) 100mg/kg 6 NJ-104-5 (Curcumin diglutarate) 100 mg/kg 7 Aspirin 100 mg/kgNJ-100-9: Turmeric oil distillation fraction which distilled mostly at105 to 118° C./high vacuum

Carrageenan-induced hind paw edema is the standard experimental model ofacute inflammation. Carrageenan is the agent of choice for testinganti-inflammatory drugs as it is not known to be antigenic and is devoidof apparent systemic effects. Moreover, the experimental model exhibitsa high degree of reproducibility (Winter, et al., Proc. Soc. Exp. Biol.Med., 1962; 111: 544-547). Carrageenan-induced edema is a biphasicresponse. The first phase is mediated through the release of histamine,serotonin and kinins whereas the second phase is related to the releaseof prostaglandin and slow reacting substances which peak at 3 hour(Vinegar R, Schreiber W, & Hugo R., J. Pharmacol. Exp. Ther., 969; 166:96-10). The increase in the paw volume following carrageenanadministration in the control and aspirin treated group corresponds withthe findings of previous workers (Jana U, Chattopadhyay R N, & Shaw B P,Indian J. Pharmacol., 1999; 31: 232-3 and Singh R K & Pandey B L. IndianJ. Physiology Pharmacol., 1996; 40: 355-8).

Results for the Carrageen-induced paw edema assay are shown in Table 20and FIG. 38.

Anti-inflammatory activity study at 100 mg/kg showed that turmeric oildistillation fraction, NJ-100-9, had 82% and 76% inhibition of paw edemaat 1 hour and 2 hours after drug administration. Curcumin diacetate wasactive at all three time points, (1 hr, 2 hr and 3 hr) by 76, 80 and 76%respectively. Fish oil inhibited paw edema by 66% at the initial timeperiods and went down to 30% by hour 3. Curcumin diglutarate and themixture of fish oil and turmeric oil showed only modest activities(below 50%) at this dose.

TABLE 20 Percent inhibition of paw edema Percent Inhibition (Std. Error)Group 1 hr 2 hr 3 hr 1 0 0 0 2 66.58 (6.46) 66.67 (9.38)   30.23 (5.37)3 82.67 (3.10) 76.39 (3.89)   54.94 (5.12) 4 48.12 (8.91) 27.78 (9.14) −6.10 (9.43) 5 76.49 (8.35) 80.56 (5.84)   76.74 (8.22) 6 49.26 (3.94)37.50 (5.11)   20.05 (8.34) 7 55.44 79.16 75.29

Results of the tail flick assay are shown in Table 21 and FIG. 40. Theanalgesic activity study showed that the combination of NJ-104-3, fishoil and the distillation fraction of turmeric oil (NJ-100-9) (combinedat 50:50 ratio), at 100 mg/kg had strong activity during the initialtime periods of 30, 60 and 90 minutes. The drugs, fish oil (NJ-104-1)and turmeric oil fraction (NJ-104-2) by themselves, were considerablyless active then the combination (NJ-104-3). Curcumin diglutarate showedstrong activity at 90 and 180 minutes. The activities of fish oil(NJ-104-1), turmeric oil (NJ-104-2) and curcumin diglutarate (NJ-104-5)were stronger than that of aspirin at 90 minutes at this dose.

TABLE 21 Tail flick latency time (seconds) in various groups Groups min30 min 60 min 90 min 120 min 180 min 1 2.4625 3.475 3.3625 3.325 2.46253.475 2 3.5375 3.875 3.8375 4.1375 3.5375 3.875 3 3.3875 3.5625 3.63.775 3.3875 3.5625 4 3.075 3.6 3.725 3.9875 3.075 3.6 5 3.4375 3.6753.6125 3.6375 3.4375 3.675 6 3.05 3.5625 3.6125 3.7375 3.05 3.5625

The anti-inflammatory activity was strong for the turmeric oil fractionand for curcumin acetate whereas the combination of turmeric oil andfish oil showed strong analgesic activity. Curcumin diglutarate alsoseem to have strong analgesic activity under the test conditions of thepresent study.

Example 10 Extraction and Purification of Turmeric Oil (II)

Turmeric powder 500 g was extracted with 3 L hexane to get 27.8 g ofcrude turmeric oil. This was distilled under high vacuum and threefractions were obtained as follow: (i) BR-103-1<108° C., 0.156 g; (ii)BR-103-2, 108-122° C., 14.48 g; and (iii) BR-103-3 3.91 g.

Fraction BR-103-2 was purified on two flash columns connected to eachother, with the first column containing 120 g of silica gel and thesecond column containing 200 g of silica gel. Eluted with hexanefollowed by 0.5% to 20% ethyl acetate, hexane as step gradientscollecting 50 mL fractions as shown in Table 22.

TABLE 22 Fractions obtained in purification of turmeric oil extractionFraction # Amount (g) Fraction label 1 1.33 BR-110-1 2 0.124 BR-110-2 30.071 BR-110-3 4 4.71 BR-110-4 5 0.594 BR-110-5 6 4.32 BR-110-6 7 3.62BR-110-7 8 0.507 BR-110-8 9 0.507 BR-110-9

A mixture (8.23 g) of turmeric oil distillation fraction BR-103-3(122-143° C./vac.) and column purified fraction BR-110-6 were combinedand distilled to get BR-132-4 (6.59 g).

A portion of BR-132-4 (300 mg) was purified by column chromatography onsilica gel (20 g), obtaining two TLC pure fractions, NJ-106-1 (16 mg)and NJ-106-2 (10 mg) for NMR spectra and the remaining as mixture.

Example 11 Extraction and Purification of Turmeric Oil (III)

Turmeric powder 500 g was extracted with 3 L hexane to get 27.8 g ofcrude turmeric oil. This was distilled under high vacuum and threefractions were obtained as follow: (i) BR-103-1<108° C., 0.156 g; (ii)BR-103-2, 108-122° C., 14.48 g; and (iii) BR-103-3 3.91 g.

Fraction BR-103-2 was purified on two flash columns connected to eachother, with the first column containing 120 g of silica gel and thesecond column containing 200 g of silica gel. Eluted with hexanefollowed by 0.5% to 20% ethyl acetate, hexane as step gradientscollecting 50 mL fractions as shown in Table 22.

Example 12 Synthesis of Curcumin Ether Derivatives

The curcumin ether derivatives were synthesized by the method describedin Majhi, et al., “Binding of curcumin and its long chain derivatives tothe activator binding domain of novel protein kinase C”, Bioorganic &Medicinal chemistry, 2010, 18: 1591-1598.

wherein R²=R₄=CH₃ and R₁=R₃=alkyl or R₁=H, R₃=alkyl.

Specifically, a mixture of curcumin (2.21 g), bromooctadecance (1.83mL), potassium carbonate (0.828 g) and acetone (60 mL) was refluxedovernight. The mixture was cooled, filtered and concentrated. Theresidue was washed with 100 mL of 10% ethyl acetate and hexane for 1hour, filtered and concentrated to get 2.0 grams, the residue waspurified over silica gel (40 grams) column, eluted with ethyl acetate (0to 10%), hexane to get two fractions: (i) BR-114-1, 0.14 grams TLCR_(f)=0.78 (Ethyl acetate, hexane 1:1), NMR consistent with structure;and (ii) BR-114-2, 0.50 grams TLC R_(f) 0.44 (Ethyl acetate, hexane1:1), NMR consistent with structure.

Example 13 Extraction of Turmeric Oil

Hexane extract (27.7 g) obtained from 500 g of turmeric powder in 3liters of hexane was distilled to obtain a distillate at 90°-105° C.,0.50 g (NJ-100-3) and a distillate at 105°-118° C., 15.5 g (NJ-100-9).

Example 14 Evaluation of Anti-Cancer Activity of Less-Purified TurmericOil Extract Fractions

The following compounds and combinations were used in this experiment: amix of 10 mg of NJ-100-3 and mg of Paclitaxel (NJ-102-2), a mix of 10 mgof NJ-100-9 and mg Paclitaxel (NJ-102-3), and Pacitaxel (NJ-102-4).

Determination of EC₅₀ of Testing Compounds:

Cytotoxicity of the compounds on three different cancer cell lines andone normal cell line was tested. Four human cell lines including bothcancer cell lines (SK-BR-3, PANC-1, PC3) and normal cell line (WI38)were initially employed. The cells (20,000 cells/well) were plated ontoa 96-well plate for 24 hours before treatment. The compounds werediluted in the medium and added to the cells at different concentrationsfor additional 48 hours from 0.04 ng/ml to 400 ng/ml. MTT assay was thenperformed. EC₅₀ values are summarized in Table 23.

TABLE 23 EC₅₀ values of compounds for various cell lines Cell lineNJ-102-2 NJ-102-3 NJ-102-4 PANC-1 7.466 ng/ml 22.53 ng/ml 32.79 ng/mlPC3 16.13 ng/ml 28.11 ng/ml 8.511 ng/ml SK-BR-3 28.08 ng/ml 26.92 ng/ml36.07 ng/ml WI38 13.91 ng/ml 16.24 ng/ml 37.91 ng/ml

Determination of GI₅₀ (50% Inhibition of Cell Growth):

Since the compounds displayed inhibitory effect on cell proliferation, aGI₅₀ experiment was performed. The cells were plated onto a 96 wellplate and treated with different concentrations of the compounds from0.04 ng/ml to 400 ng/ml for 72 hours followed by MTT assay. GI₅₀ valuesare summarized in Table 24.

TABLE 24 GI₅₀ values of compounds for various cell lines Cell lineNJ-102-2 NJ-102-3 NJ-102-4 PANC-1  2.69 ng/ml 2.599 ng/ml 5.456 ng/mlPC3  5.01 ng/ml  1.55 ng/ml  0.05 ng/ml SK-BR-3  2.72 ng/ml  0.41 ng/ml 0.78 ng/ml WI38 15.74 ng/ml 14.68 ng/ml  5.60 ng/ml

Example 15 Characterization of Turmeric Oil Components

HPLC:

Hewlet Packard, model HP 1090 series II, column X-terra C-18 5 mm,4.6×150 mm, Solvent A: acetonitrile/water 85-15, Solvent B:acetonitrile, methanol, water, phosphoric acid, 40-10-49-1.

NMR:

Brucker (400 MHz) (solvent CDCl₃ unless specified otherwise).

GC MS results were obtained on a Waters GCT Premier coupled with anAgilent 6890N GC. GC separation from a splitless injection was performedon a J&W Scientific DB-5MS column, 30 m×0.25 mm ID×25 micron filmthickness, with He carrier gas at 1 ml/min. The temperature gradient wasas follows: 50 C for 2 min, 25 C/min to 250 C, and 250 C for 4 min. Ionswere generated via electron ionization, and mass spectra were recordedat a resolving power of 6,000.

LCMS:

LCMS results were acquired with a Bruker MicrOTOF-QII quadrupoletime-of-flight mass spectrometer coupled with a Dionex Ultimate 3000RSLC ultrahigh pressure LC. LC separation was performed on a DionexAcclaim RSLC 120 C18 2.1 mm ID×100 mm column with 2.2 um particles and120 angstrom pore size. The binary gradient consisted of solvent A(water with 0.1% formic acid) and solvent B (acetonitrile with 0.1%formic acid) at 0.5 ml/min initially at 90:10 (A:B) for 2 min ramped to0:100 (A:B) at 12 min then held until 14 min. Ions were generated viaelectrospray ionization, and mass spectra were recorded at a resolvingpower of 10,000.

Separation and Characterization of Turmeric Oil Fractions:

Turmeric powder (500 g) was extracted with hexane to get 27.80 g of thecrude extract. This was distilled under high vacuum to obtain a fraction(14.48 g) that distilled at 108-122° C. This was further purified withsilica gel column chromatography using step gradient of hexane (100 to95%) and ethyl acetate (0 to 5%). This gave several column fractions andthe following were selected for the current study, CF-1, 1.33 g, CF-2,0.071 g, CF-3, 4.71 g, and CF-4, 0.594 g. CF-1 (0.50 g) was furtherpurified by a silica gel column eluting with hexane to give 0.25 g ofCF-5. HPLC and mass spectral studies for these compounds indicated thatthey were sesquiterpenes (FIG. 49).

CF-1: HPLC (solvent A), RT (area), 5.67 min (8%), 7.88 min (48%) and11.28 (43%). GCMS, RT (m/z) 9.35 min (202.2) and 9.75 min (204.2). NMR(see structural identification in CF-5).

CF-2: HPLC (solvent A), RT 3.00 min (63%), 4.20 min (11.8%), 5.02 min(19.37%), GCMS, RT at 7.54 and 9.03 minutes and mass peak at 218.1664corresponding to molecular formula, C₁₅H₂₂O. NMR, δ, 0.83-0.95 (m, 4H),1.14-1.23 (m, 4H), 1.53-2.33 (m, 26H), 2.72 (q, 1H), 3.80 (bs, 0.65H),5.20 (bs, 0.58H), 5.38 (bs, 1H), 5.98-6.1 (m, 1.8H) (unidentifiedmixture of turmerone isomers).

CF-3: HPLC (solvent A), RT, (2.87 min, 48.38%), 4.08 min (48.46%), GCMS,RT 11.08 min (m/z 216.1) 11.46 (m/z 218.2). The GC-mass spectralfragmentation peak confirms the structures, β-turmerone/curlone,α-turmerone, ar-turmerone (see supplementary information). NMR δ 0.75(m), 0.0.88 (d, J=6.5 Hz), 0.89 (d, J=6.5 Hz), 0.95 (m), 1.07 (m), 1.24(d, J=7 Hz), 1.38 (m), 1.6 (s=water), 1.64 (bs), 1.72 (q, J=1.8 Hz),1.75 (m), 1.86 (d, J=1.3 Hz), 1.88 (d, J=1.5 Hz), 1.89 (d, J=1.5 Hz),1.97-2.35 (m), 2.14 (d, J=1.1 Hz), 2.15 (d, J=1.1 Hz), 2.4-2.5 (m), 2.51(dd, J=14.6, 4 Hz), 2.61 (dd, J=15.7, 6.0 Hz), 2.68 (dd, J=15.7, 6.0Hz), 3.1 (m), 3.14 (d, J=1.2 Hz), 3.28 (m), 4.75 (bs), 4.77 (bs), 4.82(br m), 4.86 (q, J=1.2 Hz), 4.95 (br m), 5.38 (br m), 5.43 (bs), 5.64(dd, J=10, 3.2 Hz), 5.69 (br m), 5.8 (ddd, J=9.8, 2.4, 1.5 Hz), 6.02 (t,J=1.4 Hz), 6.06 (m), 6.14 (t, J=1.3 Hz), 6.17 (dd, J=10, 2.5 Hz), 7.1(br d, J=1.7 Hz). CF-2 was analyzed by ¹H NMR and determined to be amixture of β-turmerone/curlone (53%), α-turmerone (36%), ar-turmerone(4%), and an unidentified components (7%) by comparison with publishedNMR data. Percentages were calculated from relative integration valuesof the resolved peaks of each as a function of the whole sampleincluding the unidentified component(s).

CF-4: HPLC (solvent B), (RT 1.96 min, 84%), 3.26 min (15%). GCMS, RT8.45 min, m/z 216.1493, corresponds to molecular formula, C₁₅H₂₀O. NMR δ1.24 (3H, d, J=7 Hz), 1.6 (s=water), 1.85 (3H, d, J=1.3 Hz), 2.11 (3H,s), 2.3 (3H, s), 2.61 (1H, dd, J=15.7, 8.3 Hz), 2.68 (1H, dd, J=15.6,6.1 Hz), 3.28 (1H, m), 6.02 (1H, t, J=1.4 Hz), 7.08 (4H, br d, J=1.7Hz), (corresponds to ar-turmerone).

CF-5: HPLC, RT 5.64 min (3.88%), 7.93 min (49.46%), 11.27 min (46.62%),GCMS, RT 10.15 min (m/z 202, 145, 132, 119, 105, 91), RT 10.31 min (m/z204, 161, 119, 109, 93), RT 10.43 min (m/z 204, 161, 133, 120, 109, 93).The GC-MS and mass spectral fragmentation peaks confirmed the structures(data not shown). NMR δ 0.86 (d, J=6.9 Hz), 0.88 (d, J=6.8 Hz), 1.19(m), 1.23 (d, J=6.9 Hz), 1.41 (m), 1.5-1.7 (br m), 1.62 (s), 1.68 (s),1.7 (s), 1.73 (m), 1.83-2.18 (br m), 2.23 (m), 2.29 (m), 2.44 (dt,J=14.9, 4.3 Hz), 2.67 (q, J=7.2 Hz), 4.75 (br s), 5.1 (m), 5.12 (br t,J=7.1 Hz), 5.46 (br s), 5.65 (dd, J=9.9, 3.1 Hz), 5.69 (d, J=9.7 Hz),5.78 (dt, J=9.6, 2.0 Hz), 6.16 (dd, J=10, 2.7 Hz), 7.09 (m). CF-5 wasanalyzed by ¹H and 1D TOCSY NMR and determined to be a mixture ofβ-sesquiphellandrene (50.3%), 7-epi-zingiberene (30.7%) and ar-curcumene(19%) by comparison with published NMR data (Breedan, D. C. & Coates, R.M. Tetrahedron, 1994, 50, 11123; McBrien et al., J. Chem Ecol., 2002,28, 1797; and Takigawa et al., Appl. Environ. Microbio. 1993, 58, 1336).Percentages were calculated from relative integration values of theresolved peaks of each of the 3 major components. Comparison of 1H NMRshifts for various components of turmeric oil fractions are shown inTables 25-28.

TABLE 25 Comparison of 1H NMR Shifts for α-turmerone andβ-turmerone/curlone. ¹β-turmerone/curlone ²β-turmerone ¹α-turmerone²α-turmerone (δ) (δ) (δ) (δ) 6.17 (1H, dd, J = 10, 2.5 Hz) 6.16 (1H, dd,J = 10, 2.3 Hz 6.06 (1H, t, J = 1.3 Hz) 6.06 (1H, t, J = 1.3 Hz) 6.06(1H, m) 6.07 (1H, dd, J = 1.2, 1.0 Hz) 5.78 (1H, ddd, J = 9.8, 2.4, 1.5Hz)  5.8 (1H, ddd, J = 9.6, 1.9, 1.7 Hz) 5.68 (1H, m) 5.67 (1H, br d, J= 10.7 Hz) 5.64 (1H, dd, J = 10, 3.2 Hz) 5.64 (1H, dd, J = 9.6, 3.1 Hz)4.77 (1H, br s) 4.77 (1H, br s) 5.43 (1H, br s) 5.43 (1H, br s) 4.75(1H, br s) 4.75 (1H, br s)  2.5 (1H, dd, J = 14.6, 4 Hz)  2.5 (1H, dd, J= 14.5, 3.9 Hz)  2.5-2.4 (2H, m)  2.5-2.4 (2H, m) 2.3-2.0 (5H, m)2.3-2.0 (5H, m) 2.35-2.15 (3H, m) 2.35-2.15 (3H, m) 2.14 (3H, d, 1.1 Hz)2.14 (3H, d, J = 1.3 Hz) 2.14 (3H, d, J = 1.1 Hz) 2.14 (3H, d, J = 1.0Hz) 1.88 (3H, d, J = 1.5 Hz) 1.88 (3H, d, J = 1.3 Hz) 2.04 (1H, m) 2.03(1H, m) 1.71 (3H, d, J = 1.7 Hz) 1.71 (3H, d, J = 1.7 Hz) 1.88 (3H, d, J= 1.5 Hz) 1.88 (3H, d, J = 1.2 Hz) 0.88 (3H, d, J = 6.5 Hz) 0.88 (3H, d,J = 6.5 Hz) 1.64 (1H, m) 1.64 (1H, m) 1.38 (1H, m) 1.38 (1H, m) 0.89(3H, d, J = 6.6 Hz) 0.88 (3H, d, J = 6.3 Hz) ¹NMR shifts are referencedto CDCl3 (δ 7.27) at 300K. ²NMR shifts from (Biol. Pharm. Bull. 26(8)1135-1143 (2003).

TABLE 26 Comparison of ¹H NMR Shifts for ar-turmerone ¹ar-turmerone²ar-turmerone Unidentified Resonances (δ) (δ) (δ)  7.1 (4H, br d, J =1.7 Hz)  7.1 (4H, br d, J = 1.7 Hz) 6.14 (t, J = 1.3 Hz) 6.02 (1H,br t,J = 1.4 Hz) 6.02 (1H, t, J = 1.2 Hz) 5.38 (br) 3.28 (1H,br m) 3.28 (1H,ddd, J = 8.4, 6.7, 6.0 Hz) 4.98 (t, J = 1.2 Hz) 2.68 (1H, dd, J = 15.6,6.1 Hz) 2.68 (1H, dd, J = 15.7, 6.0 Hz) 4.95 (br m) 2.61 (1H, dd, 15.7,8.3 Hz )  2.6 (1H, dd, J = 15.7, 8.4 Hz) 4.86 (q, J = 1.2 Hz)  2.3 (3H,s)  2.3 (3H,s) 4.82 (br m) 2.11 (3H,d, J = 1.3 Hz) 2.10 (3H, d, J = 1.2Hz) 3.14 (d, 1.2 Hz) 1.85 (3H, d, J = 1.3 Hz) 1.85 (3H, d, J = 1.2 Hz) 3.1 (m) 1.24 (3H, d, J = 7 Hz) 1.24 (3H, d, J = 6.8 Hz) 1.98 (m) 1.92(m) 1.75 (m) 1.26 (br) 1.07 (m) 0.95 (m) 0.75 (m) ¹NMR shifts arereferenced to CDCl3 (δ 7.27) at 300K. ²NMR shifts from (Biol. Pharm.Bull. 26(8) 1135-1143 (2003).

TABLE 27 Comparison of ¹H NMR Shifts for β-sesquiphellandrene andzingiberene. ¹β-sesquiphellandrene ²β-sesquiphellandrene¹7-epi-zingiberene ³7-epi-zingiberene (δ) (δ) (δ) (δ) 6.16 (1H, dd, J =10, 2.7 Hz) 6.16 (1H, dd, J = 9.9, 2.5 Hz) 5.78 (1H, dt, J = 9.6, 2.0Hz) 5.78 (1H, dt, J = 9.8, 2.0 Hz) 5.69 (1H, d, 9.7 Hz) 5.70 (d, 1H, JD9.7 Hz) 5.65 (1H, dd, J = 9.9, 3.1 Hz) 5.64 (1H, dd, J = 9.8, 2.9 Hz)5.12 (1H, m) 5.12 (1H, m) 5.46 (1H, br s) 5.45 (1H, br s) 4.75 (2H, brs) 4.75 (2H, br s) 5.12 (1H, br t, J = 7.1 Hz) 5.11 (1H, br t, J = 7.6Hz) 2.44 (1H, dt, J = 14.9, 4.3 Hz) 2.44 (1H, dt, J = 16.5, 4.6 Hz) 2.27(1H, br m) 2.28 (1H, m) 2.29 (1H, m)  2.3 (1H, m) 2.04 (2H, m) 2.04 (2H,m) 2.23 (1H, m) 2.23 (1H, m) 2.01 (2H, m) 2.03 (2H, m) 2.01 (2H, m)  2.0(2H, m)  1.7 (3H, br s) 1.72 (3H, d, J = 1.7 Hz) 1.73 (1H, m) 1.74(1H,m) 1.68 (3H, br s) 1.68 (3H, s) 1.70 (3H, s) 1.70 (3H, s) 1.62 (3H,s) 1.61 (3H, s) 1.62 (3H, s) 1.62 (3H, s) 1.19 (1H, m) 1.18 (1H, m) 1.56(1H, m) 1.57 (1H, m) 0.88 (3H, d, J = 6.8 Hz) 0.87 (3H, d, J = 6.8 Hz)1.41 (2H, m) 1.40 (2H, m) 1.19 (1H, m) 1.22 (1H, m) 0.86 (3H, d, J = 6.9Hz) 0.86 (3H, d, J = 8 Hz) ¹NMR shifts are referenced to CDCl3 (δ 7.24)at 300K. ²NMR shifts from Biol. Pharm. Bull. 26(8) 1135-1143 (2003).³NMR shifts from Tetrahedron Vol. 50, No. 38, pp. 11123-11132. 1994.

TABLE 28 Comparison of 1H NMR Shifts for α-curcumene. ¹α-curcumene²α-curcumene (δ) (δ) 7.09 (4H, br d, J = 4 Hz) 7.05 (4H, br s)  5.1 (1H,br) 5.09 (1H, t, J = 7.7 Hz) 2.67 (1H, q, J = 7.2 Hz) 2.63 (1H, m) 1.89(2H, br m) 1.9-1.8 (2H, m) 1.7-1.5 (2H, m) 1.7-1.5 (2H, m) 1.68 (3H, s)1.66 (3H, s) 1.54 (3H, s) 1.51 (3H, s) 1.23 (3H, d, J = 6.9 Hz) 1.21(3H, d, J = 6.9 Hz)

All patents and other publications identified in the specification areexpressly incorporated herein by reference for all purposes. Thesepublications are provided solely for their disclosure prior to thefiling date of the present application. Nothing in this regard should beconstrued as an admission that the inventors are not entitled toantedate such disclosure by virtue of prior invention or for any otherreason. All statements as to the date or representation as to thecontents of these documents is based on the information available to theapplicants and does not constitute any admission as to the correctnessof the dates or contents of these documents.

1. A process for obtaining an turmeric oil extract, the processcomprising distilling turmeric oil under high vacuum distillation andcollecting a distillate at 70-100° C., 105-118° C. or at 100-130° C. 2.The turmeric oil extract of claim 1, wherein the extract hasanti-inflammatory, anti-cancer and/or analgesic activity.
 3. Theturmeric oil extract of claim 2, wherein the extract has anti-canceractivity against pancreatic, breast or prostate cancer.
 4. A compositioncomprising turmeric oil or a turmeric oil extract and a compoundselected from the group consisting of an anti-cancer compound, ananti-inflammatory agent, curcumin, a curcumin derivative, fish oil, fishoil extract, aspirin, a salicylic acid conjugate, a curcumin etherderivative or a combinations thereof, wherein the salicylic acidconjugate is of formula (II):

wherein: R⁸ is a carbohydrate; R⁹ is H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl, oroptionally substituted acyl; and analogs, derivatives, isomers,prodrugs, and pharmaceutically acceptable salts thereof, wherein thecurcumin ether derivative is of formula (III):

wherein: (i) R₁, R₂, R₃, and R₄ are independently H, alkyl, acyl,trialkylsilyl (—Si(alkyl)₃), aryl dialkylsilyl (—Si(alkyl)₂aryl),diarylalkylsilyl (—Si(aryl)₂alkyl), or triarylsilyl (—Si(aryl)₃); or(ii) R₂ and R₄ are both CH₃, and R₁ and R₃ are both alkyl or one of R₁and R₃ is H and the other is alkyl, and wherein the curcumin derivativeis of formula (I):

wherein: R¹ and R² are independently H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl, apeptide, —C(O)R³, —C(O)OR³, or —C(O)NR³R³, provided that at least one ofR¹ and R² is not H, or both of R¹ and R² are not octyl or hexadecyl, orone of R¹ or R² is not H and the other is not octyl or hexadecyl; R³ isindependently for each occurrence H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl; or R¹and R² are linking groups and at least one of R¹ and R² is a linkinggroup conjugated with a glyceryl lipid; and analogs, derivatives,isomers, prodrugs, and pharmaceutically acceptable salts thereof.
 5. Thecomposition of claim 4, wherein R⁸ is selected from the group consistingof glucose, glyceraldehydes, erythrose, threose, ribulose, xylulose,ribose, arabinose, deoxyribose, xylose, lyxose, psicose, fructose,sorbose, tagatose, allose, altrose, mannose, gulose, idose, galactose,talose, fucose, fuculose, rhamnose, sedoheptulose, octose, nonose(Neuraminic acid), and the like.
 6. The composition of claim 4, whereinat least one of R¹ and R² of Formula (I) is selected from the groupconsisting of acetyl, myristoleoyl, palmitoleoyl, sapienoyl, oleoyl,linoleoyl, α-linoleoyl, α-linolenoyl, γ-linolenoyl, arcchidionoyl,eicosapentaenoyl, erucoyl, docosahexaenoyl, lauroyl, myrsitoyl,palmitoyl, stearoyl, arachidoyl, behenoyl, lignoceroyl, certoyl and anycombinations thereof.
 7. The composition of claim 4, wherein at leastone of R¹ and R² of Formula (I) is —C(O)R³ and R³ is an optionallysubstituted aryl.
 8. The curcumin derivative of claim 4, wherein thecurcumin derivative is selected from the group consisting ofdi(acetylsalicyloyl)-curcumin monoacetylsalicyloyl-curcumin,diacetyl-curcumin, monoacetyl-curcumin, diaglutaroyl-curcumin,monoglutaroyl-curcumin, di-gluocose-glutaroyl-curcumin,mono-gluocose-glutaroyl-curcumin, monolinoleol-curcumnin,di-linoleoyl-curcumin and peptide-curcumin conjugates.
 9. Thecomposition of claim 4, wherein the composition comprises turmeric oilextract and paclitaxel; turmeric oil extract and aspirin; turmeric oilextract and a curcumin derivative, turmeric oil extract and a salicylicacid conjugate; turmeric oil extract and curcumin; turmeric oil extractand di(acetylsalicyloyl)-curcumin; turmeric oil extract andmonoacetylsalicyloyl-curcumin; turmeric oil extract anddiacetyl-curcumin; turmeric oil extract and monoacetyl-curcumin;turmeric oil extract and diaglutaroyl-curcumin; turmeric oil extract andmonoglutaroyl-curcumin; turmeric oil extract anddi-gluocose-glutaroyl-curcumin; turmeric oil extract andmono-gluocose-glutaroyl-curcumin; turmeric oil extract andmonolinoleol-curcumnin; turmeric oil extract and di-linoleoyl-curcumin;turmeric oil extract and an anticancer agent; turmeric oil extract andan anti-inflammatory agent; turmeric oil extract and fish oil; turmericoil extract and fish oil extract; turmeric oil extract, an anti-canceragent and an anti-inflammatory agent; turmeric oil extract, ananti-cancer agent and a curcumin derivative; turmeric oil extract, ananti-cancer agent and a salicylic acid conjugate; turmeric oil extract,an anti-inflammatory agent and a curcumin derivative; turmeric oilextract, an anti-inflammatory agent and a salicylic acid conjugate; orturmeric oil extract, a curcumin derivative and a salicylic acidconjugate.
 10. The composition of claim 9, wherein the composition hasenhanced anti-cancer activity.
 11. The composition of claim 4, whereinthe linking group is a glutaryl group, succinoyl group, or ether group.12. The composition of claim 4, wherein the glyceryl lipid comprises oneor two lipids conjugated with glycerol.
 13. The composition of claim 4,wherein the lipid is a fatty acid or fatty alkyl group.
 14. Thecomposition of claim 4, wherein R¹ and R² are glutaric acid and at leastone of R¹ and R² is a glutarate conjugated with a glyceryl lipid.
 15. Amethod comprising: administering a therapeutically effective amount of:a. a turmeric oil extract, b. a composition comprising: (a) turmeric oilor a turmeric oil extract, and (b) a compound selected from the groupconsisting of an anti-cancer compound, an anti-inflammatory agent, fishoil, fish oil extract, aspirin, a salicylic acid conjugate, a curcuminether derivative and any combinations thereof, c. a salicylic acidconjugate, or d. any combinations thereof to a subject in need thereof,wherein the salicylic acid conjugate is of formula (II):

wherein R⁸ is a carbohydrate; R⁹ is H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl, oroptionally substituted acyl; and analogs, derivatives, isomers,prodrugs, and pharmaceutically acceptable salts thereof, wherein thecurcumin ether derivative is of formula (III):

wherein: (i) R₁, R₂, R₃, and R₄ are independently H, alkyl, acyl,trialkylsilyl (—Si(alkyl)₃), aryl dialkylsilyl (—Si(alkyl)₂aryl),diarylalkylsilyl (—Si(aryl)₂alkyl), or triarylsilyl (—Si(aryl)₃); or(ii) R₂ and R₄ are both CH₃, and R₁ and R₃ are both alkyl or one of R₁and R₃ is H and the other is alkyl, and wherein the curcumin derivativeis of formula (I):

wherein: R¹ and R² are independently H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl, apeptide, —C(O)R³, —C(O)OR³, or —C(O)NR³R³, provided that at least one ofR¹ and R² is not H, or both of R¹ and R² are not octyl or hexadecyl, orone of R¹ or R² is not H and the other is not octyl or hexadecyl; R³ isindependently for each occurrence H, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl; or R¹and R² are linking groups and at least one of R¹ and R² is a linkinggroup conjugated with a glyceryl lipid; and analogs, derivatives,isomers, prodrugs, and pharmaceutically acceptable salts thereof. 16.The method of claim 15, wherein the linking group is a glutaryl group,succinoyl group, or ether group.
 17. The method of claim 15, wherein thelipid is a fatty acid or fatty alkyl group.
 18. The method of claim 15,wherein R¹ and R² are glutaric acid and at least one of R¹ and R² is aglutarate conjugated with a glyceryl lipid.
 19. The method of claim 15,wherein the subject is in need of treating inflammation or aninflammatory disease or condition.
 20. The method of claim 15, whereinthe subject is in need of treating cancer or a metastasis.